IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. Exp. Cell Res. (0014-4827) 2008 imex curation Lorkovic ZJ., Hilscher J., Barta A. The authors inform us that "because all SR proteins are RNA binding proteins, protein extracts used for pull-downs and immunoprecipitations were treated with RNase A to avoid indirect, RNA-mediated interactions". zdravko.lorkovic@meduniwien.ac.at 14-Nov-2008: Contacted by JYOTI. orchard Only protein-protein interactions tobac-leaf Nicotiana tabacum (tobacco) leaf leaf leaf anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lorkovic ZJ., Hilscher J., Barta A. The authors inform us that "because all SR proteins are RNA binding proteins, protein extracts used for pull-downs and immunoprecipitations were treated with RNase A to avoid indirect, RNA-mediated interactions". zdravko.lorkovic@meduniwien.ac.at Accepted 2008-NOV-16 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Exp. Cell Res. (0014-4827) 2008 orchard Only protein-protein interactions imex curation Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. Exp. Cell Res. (0014-4827) 2008 imex curation Lorkovic ZJ., Hilscher J., Barta A. The authors inform us that "because all SR proteins are RNA binding proteins, protein extracts used for pull-downs and immunoprecipitations were treated with RNase A to avoid indirect, RNA-mediated interactions". zdravko.lorkovic@meduniwien.ac.at 14-Nov-2008: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Lorkovic ZJ., Hilscher J., Barta A. The authors inform us that "because all SR proteins are RNA binding proteins, protein extracts used for pull-downs and immunoprecipitations were treated with RNase A to avoid indirect, RNA-mediated interactions". zdravko.lorkovic@meduniwien.ac.at Accepted 2008-NOV-16 by ORCHARD. 14-Nov-2008: Contacted by JYOTI. Exp. Cell Res. (0014-4827) 2008 orchard Only protein-protein interactions imex curation Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. Co-localisation studies of Arabidopsis SR splicing factors reveal different types of speckles in plant cell nuclei. Exp. Cell Res. (0014-4827) 2008 imex curation Lorkovic ZJ., Hilscher J., Barta A. The authors inform us that "because all SR proteins are RNA binding proteins, protein extracts used for pull-downs and immunoprecipitations were treated with RNase A to avoid indirect, RNA-mediated interactions". zdravko.lorkovic@meduniwien.ac.at 14-Nov-2008: Contacted by JYOTI. orchard Only protein-protein interactions tobac-leaf Nicotiana tabacum (tobacco) leaf leaf leaf confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Lorkovic ZJ., Hilscher J., Barta A. The authors inform us that "because all SR proteins are RNA binding proteins, protein extracts used for pull-downs and immunoprecipitations were treated with RNase A to avoid indirect, RNA-mediated interactions". zdravko.lorkovic@meduniwien.ac.at Accepted 2008-NOV-16 by ORCHARD. Exp. Cell Res. (0014-4827) 2008 orchard Only protein-protein interactions imex curation src28_arath Serine/arginine-rich SC35-like splicing factor SCL28 SCL28 28 kDa SC35-like splicing factor SC35-like splicing factor 28 Serine/arginine-rich splicing factor 28 F17K4.60 At5g18810 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MARARSRSRSYSPRPRDRSPPRERKGYDDNRLRERPSSRDHESSGPSGLLIRNLPLDARPNDLRDSFERFGPLKDIYLPRNYYTGEPRGFGFVKYRYAEDAAEAMKRMNHKVIGGREIAIVFAEENRKTPQEMRTTNGTSGRHGDYKRTSHRSPRRRYRSHSRSRSPPRRESRHSKVREDDLYSPRRRSRSISRSPLPRNEREYKSRNCRSPREERVLTPIRSRCLSRSRSRSLSR 1328AAD75AF1713B srs31_arath Serine/arginine-rich splicing factor RS31 RS31 RSP31 F21F14.30 At3g61860 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MRPVFVGNFEYETRQSDLERLFDKYGRVDRVDMKSGYAFVYFEDERDAEDAIRKLDNFPFGYEKRRLSVEWAKGERGRPRGDAKAPSNLKPTKTLFVINFDPIRTKEHDIEKHFEPYGKVTNVRIRRNFSFVQFETQEDATKALEATQRSKILDRVVSVEYALKDDDERDDRNGGRSPRRSLSPVYRRRPSPDYGRRPSPGQGRRPSPDYGRARSPEYDRYKGPAAYERRRSPDYGRRSSDYGRQRSPGYDRYRSRSPVPRGRP 445FD2E4A5E0C203 src30_arath Serine/arginine-rich SC35-like splicing factor SCL30 SCL30 SC35-like splicing factor 30 Serine/arginine-rich splicing factor 30 T22E16.120 At3g55460 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MRRYSPPYYSPPRRGYGGRGRSPPPPPPRRGYGGGGGGGGRRGSSHGSLLVRNIPLDCRPEELREPFERFGPVRDVYIPRDYYSGQPRGFAFVEFVDAYDAGEAQRSMNRRSFAGREITVVVASESRKRPEEMRVKTRTRSREPSGSRDRSHGRSRSRSISRSRSPRRPSDSRSRYRSRSYSPAPRRRGGPPRGEEDENYSRRSYSPGYEGAAAAAPDRDRNGDNEIREKPGYEAEDRRRGGRAVSRSPSGSRSRSVEVSPR D9D602E6F42EEDBB sr30_arath Serine/arginine-rich splicing factor SR30 SR30 SF2/ASF-like splicing modulator Srp30 SRP30 Serine-arginine rich RNA binding protein 30 T12M4.19 At1g09140 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSRWNRTIYVGNLPGDIRKCEVEDLFYKYGPIVDIDLKIPPRPPGYAFVEFEDPRDADDAIYGRDGYDFDGCRLRVEIAHGGRRFSPSVDRYSSSYSASRAPSRRSDYRVLVTGLPPSASWQDLKDHMRKAGDVCFSEVFPDRKGMSGVVDYSNYDDMKYAIRKLDATEFRNAFSSAYIRVREYESRSVSRSPDDSKSYRSRSRSRGPSCSYSSKSRSVSPARSISPRSRPLSRSRSLYSSVSRSQSRSKSRSRSRSNSPVSPVISG 73BDC3534A8F9AC4 rsz22_arath Serine/arginine-rich splicing factor RSZ22 RSZ22 RS-containing zinc finger protein 22 RSZP22 SRZ22 F28M20.230 At4g31580 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSRVYVGNLDPRVTERELEDEFRAFGVVRSVWVARRPPGYAFLDFEDPRDARDAIRALDGKNGWRVEQSHNRGERGGGGRGGDRGGGGGGRGGRGGSDLKCYECGETGHFARECRNRGGTGRRRSKSRSRTPPRYRRSPSYGRRSYSPRARSPPPPRRRSPSPPPARGRSYSRSPPPYRAREEVPYANGNGLKERRRSRS BDA0E941F763D741 ru17_arath U1 small nuclear ribonucleoprotein 70 kDa RNU1 T3A5.50 At3g50670 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGDSGDPFLRNPNAAVQARAKVQNRANVLQLKLMGQSHPTGLTNNLLKLFEPRPPLEYKPPPEKRKCPPYTGMAQFVSNFAEPGDPEYAPPKPEVELPSQKRERIHKLRLEKGVEKAAEDLKKYDPNNDPNATGDPYKTLFVSRLNYESSESKIKREFESYGPIKRVHLVTDQLTNKPKGYAFIEYMHTRDMKAAYKQADGQKIDGRRVLVDVERGRTVPNWRPRRLGGGLGTSRVGGGEEIVGEQQPQGRTSQSEEPSRPREEREKSREKGKERERSRELSHEQPRERSRDRPREDKHHRDRDQGGRDRDRDSRRDRDRTRDRGDRDRRDRDRGRDRTSRDHDRDRSRKKERDYEGGEYEHEGGGRSRERDAEYRGEPEETRGYYEDDQGDTDRYSHRYDKMEEDDFRYEREYKRSKRSESREYVR 2B35E824EF35A341 src33_arath Serine/arginine-rich SC35-like splicing factor SCL33 SCL33 SC35-like splicing factor 33 SR33 Serine/arginine-rich splicing factor 33 F7A10.15 At1g55310 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MRGRSYTPSPPRGYGRRGRSPSPRGRYGGRSRDLPTSLLVRNLRHDCRQEDLRKSFEQFGPVKDIYLPRDYYTGDPRGFGFVQFMDPADAADAKHHMDGYLLLGRELTVVFAEENRKKPTEMRARERGGGRFRDRRRTPPRYYSRSRSPPPRRGRSRSRSGDYYSPPPRRHHPRSISPREERYDGRRSYSRSPASDGSRGRSLTPVRGKSRSLSPSPRRSISRSPRRSRSPSPKRNRSVSPRRSISRSPRRSRSPRRSRRSYTPEPARSRSQSPHGGQYDEDRSPSQ 8964F965DF8ACFDD sl30a_arath Serine/arginine-rich SC35-like splicing factor SCL30A SCL30A SC35-like splicing factor 30A Serine/arginine-rich splicing factor 30A K20M4.1 At3g13570 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MRGRSYTPSPPRGYGRRGRSPSPRGRFGGSRDSDLPTSLLVRNLRHDCRQEDLRRPFEQFGPVKDIYLPRDYYTGDPRGFGFIQFMDPADAAEAKHQMDGYLLLGRELTVVFAEENRKKPTEMRTRDRGGRSNRFQDRRRSPPRYSRSPPPRRGRRSRSRSRGYNSPPAKRHQSRSVSPQDRRYEKERSYSRSPPHNGSRVRSGSPGRVKSHSRSPRRSVSPRKNRSYTPEQARSQSPVPRQSRSPTPVPRGAQNGDRSPSQ A77DDEF4E9ECD7B2 rsz21_arath Serine/arginine-rich splicing factor RSZ21 RSZ21 RS-containing zinc finger protein 21 RSZP21 SRZ21 T23E23.2 At1g23860 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MTRVYVGNLDPRVTERELEDEFKAFGVLRNVWVARRPPGYAFLEFDDERDALDAISALDRKNGWRVELSHKDKGGRGGGGGRRGGIEDSKCYECGELGHFARECRRGRGSVRRRSPSPRRRRSPDYGYARRSISPRGRRSPPRRRSVTPPRRGRSYSRSPPYRGSRRDSPRRRDSPYGRRSPYANGV D8706B45A8E78F72 q9fyb1_arath sc35 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSHFGRSGPPDISDTYSLLVLNITFRTTADDLYPLFAKYGKVVDVFIPRDRRTGDSRGFAFVRYKYKDEAHKAVERLDGRVVDGREITVQFAKYGPNAEKISKGRVVEPPPKSRRSRSRSPRRSRSPRRSRSPPRRRSPRRSRSPRRRSRDDYREKDYRKRSRSRSYDRRERHEEKDRDHRRRTRSRSASPDEKRRVRGRYDNESRSHSRSLSASPARRSPRSSSPQKTSPAREVSPDKRSNERSPSPRRSLSPRSPALQKASPSKDMSPERRSNERSPSPGSPAPLRKVDAASRSQSPYAAE 05159A75E7AED8A1 sr34_arath Serine/arginine-rich-splicing factor SR34 SR34 Pre-mRNA-splicing factor SF2 SF2 SR1 SR1 protein SRP34 F22D16.16 F22D16_30 At1g02840 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MSSRSSRTVYVGNLPGDIREREVEDLFSKYGPVVQIDLKVPPRPPGYAFVEFDDARDAEDAIHGRDGYDFDGHRLRVELAHGGRRSSDDTRGSFNGGGRGGGRGRGDGGSRGPSRRSEFRVLVTGLPSSASWQDLKDHMRKGGDVCFSQVYRDARGTTGVVDYTCYEDMKYALKKLDDTEFRNAFSNGYVRVREYDSRKDSRSPSRGRSYSKSRSRSRGRSVSRSRSRSRSRSRSPKAKSSRRSPAKSTSRSPGPRSKSRSPSPRRSRSRSRSPLPSVQKEGSKSPSKPSPAKSPIHTRSPSR 6530F9CB628B8EFA rsz33_arath Serine/arginine-rich splicing factor RS2Z33 RS2Z33 RS-containing zinc finger protein 33 RSZ33 F3G5.13 At2g37340 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPRYDDRYGNTRLYVGRLSSRTRTRDLERLFSRYGRVRDVDMKRDYAFVEFGDPRDADDARHYLDGRDFDGSRITVEFSRGAPRGSRDFDSRGPPPGAGRCFNCGVDGHWARDCTAGDWKNKCYRCGERGHIERNCKNSPKKLRRSGSYSRSPVRSRSPRRRRSPSRSLSRSRSYSRSRSPVRRRERSVEERSRSPKRMDDSLSPRARDRSPVLDDEGSPKIIDGSPPPSPKLQKEVGSDRDGGSPQDNGRNSVVSPVVGAGGDSSKEDRSPVDDDYEPNRTSPRGSESP 4B0407A7E106EFA2 rszp21-rszp22 3 8 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 12 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 1B Co-expression of RSZp21-GFP and RSZp22-RFP also resulted in their complete co-localisation Co-expression of RSZp21-GFP and RSZp22-RFP also resulted in their complete co-localisation in nuclear speckles. rszp21-rszp33 3 15 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 12 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 1B Co-expression of RSZp21 and RSZp22 fused to GFP with RSZ33-RFP resulted in co-localisation of each protein pair Co-expression of RSZp21-GFP and RSZp33-RFP also resulted in their complete co-localisation in nuclear speckles. rszp22-rszp33 3 15 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 8 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 1C Co-expression of RSZp21 and RSZp22 fused to GFP with RSZ33-RFP resulted in co-localisation of each protein pair Co-expression of RSZp22-GFP and RSZp33-RFP also resulted in their complete co-localisation in nuclear speckles. sf2-rnu1 3 9 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 14 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 3B However, co-expression of SRp34-RFP with the U1 snRNP-specific protein 70K fused to GFP, another protein pair known to interact in vitro and in vivo [26,31], resulted in their precise co-localisation as reported previously Proteins colocalised within nuclear speckles. rsz33-rnu1-1 3 15 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 9 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 3C In line with the above results cells coexpressing RSZ33-RFP and U1-70K-GFP showed only partial co-localisation Proteins were shown to partially colocalise within nuclear speckles. rsz33-scl28 3 15 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization The authors state that "co-expression of RSZ33-GFP with SCL28, SCL30, and SCL30a fused to RFP resulted in their partial co-localisation", however do not show the data in the case of SCL28 / RSZ33 colocalisation. Co-expression of RSZ33-GFP with SCL28, SCL30, and SCL30a fused to RFP resulted in their partial co-localisation rsz33-scl30 3 6 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 15 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Co-expression of RSZ33-GFP with SCL28, SCL30, and SCL30a fused to RFP resulted in their partial co-localisation Figure 4A The proteins partially colocalised to nuclear speckles. rsz33-scl30a 3 15 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Co-expression of RSZ33-GFP with SCL28, SCL30, and SCL30a fused to RFP resulted in their partial co-localisation Figure 4C The proteins partially colocalised to nuclear speckles. scl28-scl30 3 4 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 5A Co-expression of protein pairs indicated in Fig. 5A clearly revealed that SCL proteins do co-localise with each other. The proteins partially colocalised to nuclear speckles. scl28-scl33 3 10 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 5A Co-expression of protein pairs indicated in Fig. 5A clearly revealed that SCL proteins do co-localise with each other. The proteins partially colocalised to nuclear speckles. sc35-scl28 3 13 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Coexpression of atSC35 with SCL proteins resulted in nearly complete co-localisation Figure 5B sc35-scl30 3 6 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 13 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Coexpression of atSC35 with SCL proteins resulted in nearly complete co-localisation Figure 5B sc35-scl30a 3 13 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Coexpression of atSC35 with SCL proteins resulted in nearly complete co-localisation Figure 5B sc35-rsz33 3 15 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 13 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 6 Therefore, co-localisation of atSC35 with RSZ33 was analysed. As evident from Fig. 6 the two proteins showed precise co-localisation in maximum intensity projections as well as in single confocal images Proteins were shown to colocalise within nuclear speckles. sc35-sf2 3 13 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 14 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 7 In contrast, cells co-expressing atSC35 and SRp34/SR1 showed only partial co-localisation which is also evident from quantification of fluorescence intensities along the three lines Proteins were shown to partially colocalise within nuclear speckles. rsp31-scl30a 3 11 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization co-expression of RSp31-GFP with SCL30a-RFP and with SRp30-RFP (another ortholog of the human SF2/ASF) also resulted in complete co-localisation of these fusion proteins Figure 7B rsp31-scl30 3 5 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization co-expression of RSp31-GFP with SCL30a-RFP and with SRp30-RFP (another ortholog of the human SF2/ASF) also resulted in complete co-localisation of these fusion proteins Figure 7B rszp21-sf2 2 12 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8A By using in vitro pull-down assay we could recapitulate interaction between RSZp21 and SRp34/SR1 scl28-scl30a-1 2 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl28-scl30-1 2 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl28 2 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl28-sf2 2 14 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl28-srp30 2 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl28-rszp21 2 12 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl33-scl28 1 10 unspecified role unspecified role bait bait c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 4 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 8C Interaction of SCL33 with SCL28 (lanes 1), SCL30 (lanes 2), SCL30a (lanes 3), and SCL33 (lanes 4). SCL33-GFP was co-expressed in tobacco protoplasts with the indicated HA-tagged proteins. Protein extracts were immunoprecipitated with anti-GFP antibody scl33-scl30 1 10 unspecified role unspecified role bait bait c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 8C Interaction of SCL33 with SCL28 (lanes 1), SCL30 (lanes 2), SCL30a (lanes 3), and SCL33 (lanes 4). SCL33-GFP was co-expressed in tobacco protoplasts with the indicated HA-tagged proteins. Protein extracts were immunoprecipitated with anti-GFP antibody scl33 1 10 unspecified role unspecified role bait bait c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 8C Interaction of SCL33 with SCL28 (lanes 1), SCL30 (lanes 2), SCL30a (lanes 3), and SCL33 (lanes 4). SCL33-GFP was co-expressed in tobacco protoplasts with the indicated HA-tagged proteins. Protein extracts were immunoprecipitated with anti-GFP antibody sc35-scl30-1 1 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role bait bait c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 8D Interaction of atSC35 with SCL30 (lanes 1), SCL30a (lanes 2), SCL33 (lanes 3), and atSC35 sc35-scl30a-1 1 13 unspecified role unspecified role bait bait c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 8D Interaction of atSC35 with SCL30 (lanes 1), SCL30a (lanes 2), SCL33 (lanes 3), and atSC35 sc35-scl33 1 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 13 unspecified role unspecified role bait bait c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 8D Interaction of atSC35 with SCL30 (lanes 1), SCL30a (lanes 2), SCL33 (lanes 3), and atSC35 scl30-scl30a-2 3 6 unspecified role unspecified role neutral component neutral component c-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 11 unspecified role unspecified role neutral component neutral component c-terminal rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range colocalization colocalization Figure 5A Co-expression of protein pairs indicated in Fig. 5A clearly revealed that SCL proteins do co-localise with each other. The proteins partially colocalised to nuclear speckles. scl30-scl28 2 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30-3 2 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30-scl30a-3 2 11 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30-rszp21 2 12 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30-srp30 2 6 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30-sf2-1 2 14 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 6 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) physical association physical association Figure 8B scl30a-scl28 2 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30a-1 2 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30a-sf2 2 14 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30a-srp30-1 2 7 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a scl30a-rszp21 2 12 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a sc30a-scl30 2 11 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8B Similarly, pull-down assays confirmed interactions of SCL28, SCL30 and SCL30a with RSZp21, SRp30, SRp34/SR1, SCL28, SCL30, and SCL30a rsz33-sf2-2 2 15 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 14 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Figure 8A no suitable source text