UniProt UniProt http://www.uniprot.org sasaki-2000-3 Yeast Krr1p physically and functionally interacts with a novel essential Kri1p, and both proteins are required for 40S ribosome biogenesis in the nucleolus. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Sasaki T., Toh-E A., Kikuchi Y. kikuchi@biol.s.u-tokyo.ac.jp Mol. Cell. Biol. 20 (7971-7979) 2000 sasaki-2000-1 Yeast Krr1p physically and functionally interacts with a novel essential Kri1p, and both proteins are required for 40S ribosome biogenesis in the nucleolus. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Sasaki T., Toh-E A., Kikuchi Y. kikuchi@biol.s.u-tokyo.ac.jp Mol. Cell. Biol. 20 (7971-7979) 2000 sasaki-2000-2 Yeast Krr1p physically and functionally interacts with a novel essential Kri1p, and both proteins are required for 40S ribosome biogenesis in the nucleolus. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Sasaki T., Toh-E A., Kikuchi Y. kikuchi@biol.s.u-tokyo.ac.jp Mol. Cell. Biol. 20 (7971-7979) 2000 fbrl_yeast rRNA 2'-O-methyltransferase fibrillarin NOP1 LOT3 YDL014W D2870 U3 small nucleolar RNA-associated protein NOP1 Histone-glutamine methyltransferase protein protein yeast Saccharomyces cerevisiae Baker's yeast MSFRPGSRGGSRGGSRGGFGGRGGSRGGARGGSRGGFGGRGGSRGGARGGSRGGFGGRGGSRGGARGGSRGGRGGAAGGARGGAKVVIEPHRHAGVYIARGKEDLLVTKNMAPGESVYGEKRISVEEPSKEDGVPPTKVEYRVWNPFRSKLAAGIMGGLDELFIAPGKKVLYLGAASGTSVSHVSDVVGPEGVVYAVEFSHRPGRELISMAKKRPNIIPIIEDARHPQKYRMLIGMVDCVFADVAQPDQARIIALNSHMFLKDQGGVVISIKANCIDSTVDAETVFAREVQKLREERIKPLEQLTLEPYERDHCIVVGRYMRSGLKK 56A8B958A7B6066E krr1_yeast KRR1 small subunit processome component KRR1 KRR-R motif-containing protein 1 Ribosomal RNA assembly protein KRR1 YCL59C YCL059C protein protein yeast Saccharomyces cerevisiae Baker's yeast MVSTHNRDKPWDTDDIDKWKIEEFKEEDNASGQPFAEESSFMTLFPKYRESYLKTIWNDVTRALDKHNIACVLDLVEGSMTVKTTRKTYDPAIILKARDLIKLLARSVPFPQAVKILQDDMACDVIKIGNFVTNKERFVKRRQRLVGPNGNTLKALELLTKCYILVQGNTVSAMGPFKGLKEVRRVVEDCMKNIHPIYHIKELMIKRELAKRPELANEDWSRFLPMFKKRNVARKKPKKIRNVEKKVYTPFPPAQLPRKVDLEIESGEYFLSKREKQMKKLNEQKEKQMEREIERQEERAKDFIAPEEEAYKPNQN 7A7F964E2C7FD056 kri1_yeast Protein KRI1 YNL308C N0388 KRR1-interacting protein 1 KRI1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPRKKSAAKRAREQAKKEAAVPATDTATIKTSETSATTVKPAIEASKSYVPSEDEEEDEEEEEEEDDYGELITDEVENGINQVLDAIKNNKTDKLLDPKVKFFEDPESAAAKLANREGKHKPIYLKDYHRMNILSGDALKEDDEEYEHATVDGKQSFVSQQREEKTQLLNEIKSAFSDEENEESSGDEDDGFLKKKEPSTKKEGKNLPDPTVNEENFLEEFVNQQAWIPKKGDKVISLDLNNNEEDDEEFEDAAEKFENAYNFRYEDPNAAEIISYARSQATLRRSDDSSRRRKREEKRKIKEQIKAEKETALQKKKTKKLNKLTDILEQLTKEYGAEINADMVKKITDTLLKNDFKEEEWDNVVAELFNEEFYQQEGKPTWNEDDEIMGDFYADADGDDQTEEGEVEKEQKEEDEEEGPKRKKSKKEEKLQKKKEKRKVNELVENALEQNKLALIEEVEKEEEERKSRSRTKEEQDLKFRYREVSPESFGLTAREIFAADDTDLNEFIGLKKFAPYRSKELRAKDKRKVMKARRLREWRKKTFKNENGLAPVEAEAGEKDEDTILIPVEKASKSKHKRGHSHKHKGHQKK 3B637571EDB05EA6 tom1_yeast E3 ubiquitin-protein ligase TOM1 TOM1 SSR2 YDR457W Temperature-dependent organization in mitotic nucleus protein 1 Suppressor of snRNA protein 2 D8035.1 HECT-type E3 ubiquitin transferase TOM1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVLFTRCEKARKEKLAAGYKPLVDYLIDCDTPTFLERIEAIQEWDRSRDDLYVWIPILDRMDGLLLKVAEKYKYKQDPKKECEVKLVEMEAHDVDYCLKMLKFTRRLLLNTENRFVYSSGDVLMYLLNCPNFTIKLAVMRILAILGERFVIAREKIVAHNIFGDHNLRKKTLKLALSLSSSVMDEDGEHFSLVDLYFDKKKVPQKWRKLRFTHYTSNDFKKSSQQKNNINETQTSIKKVTMTTQELCEHSLQQIFDKGMALLPAESWFDFSIKASVAKAFSDDSGENIDLRNIIIETKLNAIAFVNTIFSPPQVSSKLFELDPYAFNSLTDLISLSETKIPKELRTDALFTLECISLKHVWCSDIIRNLGGNISHGLLFQILRYIAKTLREATDEIDEEYNVRFFYLISNLADVKPLHESLFAAGLIPTLLEIVSIRNCPYKRTLASATHLLETFIDNSETTTEFIENDGFTMLITSVANEIDFTLAHPETWQPPKYSVVYYSISFRELAYIRSLLKLVLKLLSTDSGDRIRNLIDSPILVSLKKILENKLVFGLTLITYTLDVVQKVINSEPTIYPVLVEAGLIPYVIDNFPKLIGPSAELLSLLPDVVSAICLNPEGLKQVKEKGLINNLFDFLLDADHARILTGGDRSTEYGTDIDELARHYPDLKANIVEALCNVIRKMPSTFRNEREFLFTSPKDQKYFFHRKNEEILTDKEEHEPAYWELLDKGTMLDTFTSVLFGMSLGNGSFSQVPQHLEARDFLAIIFMENPPYEYFTSVAISNVTEVLQYLDEKYEDYAFMDVMKVLNDQLENLNDFLNSPNDRSFFLERDGENSVRSCHSKLCRLAAILNIVTNVYIDLTTLSCKRIMQIYSYFDKRGFSLIKNLKLLFQKCALEEMYIRQHMPDSVITETMPLPIVDVSGDGPPLQIYIDDPKKGDQKGKITSVKTRNTLQMRTILYTLQSNTAILFRCFLRLSHSRNMDLEHKDLTTEVHIFENVVENVIEMLKATELEGHLPYILVLLNFNTFVFTIPKASPNSTEILQTIPAYIFYQKGGYLLYLHIIRDLFTRMTKIKDLSSLDNINYIDESNGILTLSCLINALTFYNKSMQTETMENVQSIGKYYVSIDDDYNIMKALTVPIKVMALAMILDLDKSDSLFKTQSRNVPYSVFKQLLSMLKNIFTNVNIYTKELYELHWDLIFPPIKKISLFEQVGIPGDVAANYLTDTGDDLPADNSIGLFSPEQWEKYKKLIGEDKSIYYPQPMQAQYYKGCSSKELDELRDTFFNDGLPSRIFTVLPFYPKLVNAFAKTLLQIFTKYDEPTEVFAGRILDRILETDLDDPATLSSLIHLFGIFLNEKYIYQKASHLMQRFIEYLEKSLKPEHVNTPWFSKALYVYEIILAKSELPHLEELSKDVLLRYPLLSMAKVFRIPDPMKQKLFDILIRVSDISNFYSALATSRILIFYSRDELYANNIARSGILSRLLKVIGSFQKLDKINFLESSFLLLTRRCFETTENVDALIRAEINRSFTARPLGGGDDAVRELTTILEEKAHVVMRSPSQFIDVLCETARFHEFDDQGALVDYSLKRFLGEKDKNTQASSTEKSDIYERTGIMHLLLSQLMAASEKDWLSEPANSSDLPENKKAQLDPSRNPVCAYMIFLLKLLVELVSSYNQCKFEFLTFSRRNTYAERPRPRTTAINFFLYRLLDKPVGTDHDKHEAKRREVIGMLARSVIIGFLATVQDDRTTKTDVKLADPHMNFIRKFAIEAIIKAIRNATSSSKLLESNHLKLDMWFRIITSMVYVQAPYLRQLLDSNKVEADQYQLCKLVIDLGLPSVITEAMASIDLNYPFSKKIFNVAVEALNTISSTRNNFSEHFKIEDHDEVEDEVDESDKEEIPDMFKNSALGMYDVEDIEEDDDDDTSLIGDDDAMAFVDSDNGFEVVFSDEDDDMGEEDADDARSDSEENELSSEMQSSTADGTDVDYEVDDADGLIINIDQPSGDDEEMADYDANISHSSHSENEDDASMDVIEVYDDELSSGYDVDLSDYDVDESDWDSGLSSLSISDEDSESSEDEPINSTRMGDSRRRWLIAEGVELTDDSQGESEEDDRGVFRGIEHIFSNENEPLFRVHDEMRHRNHHRSINRTHFHSAMSAPSLSLLNRGRRNQSNLINPLGPTGLEQVENDISDQVTVAGSGSRPRSHHLHFSEVLVSGSFFDEPVLDGIILKSTVSRWKDIFDMFYDSKTYANCIIPTVINRLYKVSLALQKDLENKREQEKLKNKNLLFNEAKVESHNSSDAISVEQDDIQESNVTHDDHEPVYVTIQGSEVDIGGTDIDPEFMNALPDDIRADVFAQHVRERRAEARLNSDHNVHSREIDSDFLEAIPEDIREGILDTEAEEQRMFGRIGSSADVIRADDDVSNNDEEVENGLDHGNSNDRNNADPEKKKPARIYFAPLIDRAGIASLMKSVFISKPYIQREIYHELFYRLCSSKQNRNDLMNTFLFILSEGIIDQHSLEKVYNIISSRAMGHAKTTTVRQLPSDCTPLTVANQTIEILQSLIDADSRLKYFLIAEHDNLIVNKANNKSRKEALPDKKLRWPLWHLFSLLDRKLITDESVLMDLLTRILQVCTKTLAVLSTSSNGKENLSKKFHLPSFDEDDLMKILSIIMLDSCTTRVFQQTLNIIYNLSKLQGCMSIFTKHLVSLAISIMSKLKSALDGLSREVGTITTGMEINSELLQKFTLPSSDQAKLLKILTTVDFLYTHKRKEEERNVKDLQSLYDKMNGGPVWSSLSECLSQFEKSQAINTSATILLPLIESLMVVCRRSDLSQNRNTAVKYEDAKLLDFSKTRVENLFFPFTDAHKKLLNQMIRSNPKLMSGPFALLVKNPKVLDFDNKRYFFNAKLKSDNQERPKLPITVRREQVFLDSYRALFFKTNDEIKNSKLEITFKGESGVDAGGVTREWYQVLSRQMFNPDYALFLPVPSDKTTFHPNRTSGINPEHLSFFKFIGMIIGKAIRDQCFLDCHFSREVYKNILGRPVSLKDMESLDPDYYKSLVWILENDITDIIEETFSVETDDYGEHKVINLIEGGKDIIVTEANKQDYVKKVVEYKLQTSVKEQMDNFLVGFYALISKDLITIFDEQELELLISGLPDIDVDDWKNNTTYVNYTATCKEVSYFWRAVRSFDAEERAKLLQFVTGTSKVPLNGFKELSGVNGVCKFSIHRDFGSSERLPSSHTCFNQLNLPPYESYETLRGSLLLAINEGHEGFGLA 8F71F3493D70A6C9 krr1-nop1 683014 N/A 683017 unspecified role unspecified role neutral component neutral component c-terminus 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 false N/A 683018 unspecified role unspecified role neutral component neutral component c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig. 3 Both Krr1 and Nop1 localize in the nucleolus. kri1-nop1 683014 N/A 683017 unspecified role unspecified role neutral component neutral component c-terminus 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 false N/A 683019 unspecified role unspecified role neutral component neutral component c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig. 3 Both Krr1 and Nop1 localize in the nucleolus. tom1-krr1 TOM1 interacts with KRR1 in a yeast two-hybrid assay 683015 N/A 683018 unspecified role unspecified role prey prey N/A 683020 unspecified role unspecified role bait bait physical association physical association false false false No figure krr1-kri1 683016 N/A 683019 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 683018 unspecified role unspecified role bait bait P25586:p.[Phe45Leu;Leu95Ser;Arg207Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P25586:p.[Lys20Glu;Lys66Asn;Cys162Arg;Asp261Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false c-terminus ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig. 7A & 7B Interactions were performed at 26 and 37 degrees Celsius. A further decrease of the interaction was observed at 37 degrees Celsius in the case of the two mutants.