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/ http://www.ebi.ac.uk/ ashe-2004-4 LEK1 is a potential inhibitor of pocket protein-mediated cellular processes. mouse-myeloma Mus musculus myeloma cell line myeloma lymphoblast plasma cell line. coip coimmunoprecipitation co-immunoprecipitation CoIp immunoprecipitation Co-IP co-immunoprecipitation CoIp immunoprecipitation Co-IP coip western blot western blot Immuno blot Immuno blot Ashe M., Pabon-Pena L., Dees E., Price KL., Bader D. david.bader@mcmail.vanderbilt.edu J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ashe-2004-3 LEK1 is a potential inhibitor of pocket protein-mediated cellular processes. chlae-ts_cos Chlorocebus aethiops Grivet ts_cos Simian cells (cercopithecus aethiops) transformed by SV40 pull down pull down anti tag western anti tag western blot anti tag western Ashe M., Pabon-Pena L., Dees E., Price KL., Bader D. david.bader@mcmail.vanderbilt.edu J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ashe-2004-2 LEK1 is a potential inhibitor of pocket protein-mediated cellular processes. chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 coip coimmunoprecipitation co-immunoprecipitation CoIp immunoprecipitation Co-IP co-immunoprecipitation CoIp immunoprecipitation Co-IP coip western blot western blot Immuno blot Immuno blot Ashe M., Pabon-Pena L., Dees E., Price KL., Bader D. david.bader@mcmail.vanderbilt.edu J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ashe-2004-1 LEK1 is a potential inhibitor of pocket protein-mediated cellular processes. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ashe M., Pabon-Pena L., Dees E., Price KL., Bader D. david.bader@mcmail.vanderbilt.edu J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation q155p7_mouse Cenpf Lek1 protein protein mouse Mus musculus Mouse MSWALEEWKEGLPSRALQKIQELEGQLEKLKKEKQQRQFQLDSLEAALQKQKQKVEDGKTEGADLKRENQRLMEICEHLEKSRQKLSHELQVKESQVNLQESQLSSCKKQIEKLEQELKRCKSEFERSQQVAQSADVSLNPCSTPQKLFATPLTPSSTYEDLKEKYNKEVEERKRLEEEVKALHAKKVSLPVSQATMNHRDIARHQASSSVFPWQQENTPSRLSSDALKTPLRRDGSAAHFLGEEVSPNKSSMKTGRGDCSSLPGEPHSAQLLHQAKAQNQDLKSKMTELELRLQGQEKEMRSQVNKCQDLQLQLEKTKVELIEKERILNKTRDEVVRSTAQYDQAAAKCTTLEQKLKTLTEELSCHRQNAESAKRSLEQRIKEKEKELQEELSRQHQSFQALDSEYTQMKTRLTQELQQVKHLHSTLQLELEKVTSVKQQLERNLEEIRLKLSRAEQALQASQVAENELRRSSEEMKKENSLIRSQSEQRTREVCHLEEELGKVKVSLSKSQNFAEEMKAKNTSQEIMLRDLQEKLNQQENSLTLEKLKLALADLERQRNCSQDLLKKREHHIDQLNNKLNKIEKEFETLLSALELKKKECEELKEEKNQISFWKIDSEKLINQIESEKEILLGKINHLETSLKTQQVSPDSNERIRTLEMERENFTVEIKNLQSMLDSKMVEIKTQKQAYLELQQKSESSDQKHQKEIENMCLKANKLTGQVESLECKLQLLSSEVVTKDQQYQDLRMEYETLRDLLKSRGSSLVTNEDNQRSSEDNQRSSEDNQRGSLAFEQQPAVSDSFANVMGRKGSINSERSDCSVDGGRSPEHIAILQNRVTSLESSLESQNQMNSDLQMRCEELLQIKGEVEENLSKAEQIHQNFVAETNQCISKLQEDAAVHQNIVAETLATLESKEKELQLLKEKLEAQQTEVQKLNKNNCLLEGTLKELQLLSDTLSSEKKEMNSIISLSKKNIEELTQANEALKEVNEALEQEKMNLLQKHEKITSCIAEQERSIAELSDQYKQERLQLLQRCEETEAVLEDLRGNYKTAQENNAKLECMLSECTALCENRKNELEQLKETFAKEQQEFLTKLAFAEEQNRKLMLELEIEQQTVRSEITNTNKHSMSATDGLRQECLTLNEEQNEQQNEVSNLTHENEQLMELTQTKHDSYLAVEPVENSVKATEDEIGKSSSQYQMDIDTKDISLDSYKAQLVHLEALVRILEVQLDQSEEENKKLHLELQTIREELETKSSQDPQSQARTGLKDCDTAEEKYVSMLQELSASQNENAHLQCSLQTAVNKLNELGKMCDVLRVEKLQLESELNDSRTECITATSQMTAEVEKLVSEMKMLNHESALSQNELMKDTSGGEFHDKANHSSVFLTPLDSSNFCEQMTLSSKEVRVHFAELQEKFSCLQSEHKILHDQHCEVSSKMSALRSYVDTLKAENSALSMSLRTLQGDLVKEGEPAAEGGHGLPLSFCGADSPSLTNFGETSFYKDVLEQTGDTCHLSLEGNASANSCDLDEEFSSSLEEETLTEKESPPAPGRTVEGLEVLCQVYLQSLKNLEEKTESQRIMKNKEIEELEQLLSSERKELSCLRKQYLSEKEQWQQKLTSVTLEMESKLAEEKQQTKTLSLELEVARLQLQELDLSSRSLLGTDLESVVRCQNDNYDIKESEVYISETTEKTPKQDTDQTCDKDIQQDLGLETSVTESETTRLTGEGCEEQPPKTNCEAPAEDKTQDCSECISELCSSSNVLVPMDVLEDQGSIQNLQLQKDTLNENLRLLPEVEDWDKKVESLLNEIMEADSKLSLQEVQLKMKIATCIQLEKIVKDLRKEKADLSEKLESLPCNQEVCLRVERSEEDLGFNLDMGANELLSKSTKDNATNTEDNYKEKFLDMERELTRIKSEKANIEHHILSVETNLEVVQAEKLCLERDTESKQKVIIDLKEELFTVISERNRLREELDNVSKESKALDQMSKKMKEKIEELESHQRESLRHIGAVESEVKDKADLIQTLSFNVGELTKDKAHLQEQLQNLQNDSQELSLAIGELEIQIGQLNKEKESLVKESQNFQIKLTESECEKQTISKALEVALKEKGEFAVQLSSAQEEVHQLRRGIEKLSVRIEADEKKHLSAVAKLKESQRESDSLKDTVETLERELERSEENQELAILDSENLKAEVETLKAQKDEMTKSLRIFELDLVTVRTERENLAKQLQEKQSRVSELDERCSSLRRLLEEKEQARVQMEEDSKSAMLMLQMQLKELREEVAALCNDQETLKAQEQSLDQPGEEVHHLKSSIRKLKVHIDADEKKHQNILEQLKESKHHADLLKDRVENLEQELILSEKNMIFQAEKSKAEIQTLKSEIQRMAQNLQDLQLELISTRSENENLIKELKKEQERVSDLETINPSIENLLKDKEQEKVQMKEEAKITVEMLQTQLKELNETVVSLCNDQEVSKTKEQNLGSQVQTLELEKAQLLQDLGEAKNKYIIFQSSVNALTQEVEAGKQKLEKGEKEIRTLKEQLKSQEQLVCKLAQVEGEQELWQKQKLELRNVTMALEQKVQVLQSENNTLQSTYEALQNSHKSLESELGLIKLEKVALVERVSTISGKEAELQRELRDMLQKTTQLSEDYNKEKNRLTEEVEVLREELQNTKAAHLKSVNQLEKELQRAQGKIKLMLKSCRQLEGEKEMLQKELSQLEAAQQQRAGSLVDSNVDEVMTENKALKETLEEKVKEADKYLDKYCSLLISHEELEKAKEILEIEVARLKSRQSRQDLQSSPLLNSSIPGPSPNTSVSEMKSASGQNKASGKRQRSSGVWEHGKRAAPSTAETFSKKSRKSDSKSTRPAEHEQETEFEPEGLPEVVKKGFADIPTGKTSPYILRRTTMATRTSPRFATQKLVGSSPSLGKENVVESSKPTAGGSRSQKVKVVQESSADSHTAFQELPAKSLTASNIPGRNSTESPREGLRAKRAYPASSPAAGPDPTNNENCRVQ rb_mouse Retinoblastoma-associated protein Rb1 Rb-1 pRb pp105 p110-RB1 protein protein mouse Mus musculus Mouse MPPKAPRRAAAAEPPPPPPPPPREDDPAQDSGPEELPLARLEFEEIEEPEFIALCQKLKVPDHVRERAWLTWEKVSSVDGILEGYIQKKKELWGICIFIAAVDLDEMPFTFTELQKSIETSVYKFFDLLKEIDTSTKVDNAMSRLLKKYNVLCALYSKLERTCELIYLTQPSSALSTEINSMLVLKISWITFLLAKGEVLQMEDDLVISFQLMLCVVDYFIKFSPPALLREPYKTAAIPINGSPRTPRRGQNRSARIAKQLENDTRIIEVLCKEHECNIDEVKNVYFKNFIPFINSLGIVSSNGLPEVESLSKRYEEVYLKNKDLDARLFLDHDKTLQTDPIDSFETERTPRKNNPDEEANVVTPHTPVRTVMNTIQQLMVILNSASDQPSENLISYFNNCTVNPKENILKRVKDVGHIFKEKFANAVGQGCVDIGVQRYKLGVRLYYRVMESMLKSEEERLSIQNFSKLLNDNIFHMSLLACALEVVMATYSRSTLQHLDSGTDLSFPWILNVLNLKAFDFYKVIESFIKVEANLTREMIKHLERCEHRIMESLAWLSDSPLFDLIKQSKDGEGPDNLEPACPLSLPLQGNHTAADMYLSPLRSPKKRTSTTRVNSAANTETQAASAFHTQKPLKSTSLALFYKKVYRLAYLRLNTLCARLLSDHPELEHIIWTLFQHTLQNEYELMRDRHLDQIMMCSMYGICKVKNIDLKFKIIVTAYKDLPHAAQETFKRVLIREEEFDSIIVFYNSVFMQRLKTNILQYASTRPPTLSPIPHIPRSPYKFSSSPLRIPGGNIYISPLKSPYKISEGLPTPTKMTPRSRILVSIGESFGTSEKFQKINQMVCNSDRVLKRSAEGGNPPKPLKKLRFDIEGADEADGSKHLPAESKFQQKLAEMTSTRTRMQKQRMNESKDVSNKEEK rbl2_mouse Retinoblastoma-like protein 2 Rbl2 Retinoblastoma-related protein 2 130 kDa retinoblastoma-associated protein pRb2 protein protein mouse Mus musculus Mouse MASGGNQSPPPPPAAAASSEEEEEDGDAADRAQPAGSPSHQIQQRFEELCSRLNMDEAARAEAWSSYRSMSESYTLEGNDLHWLACALYVACRKSVPTVSKGTAEGNYVSLTRILRCSEQSLIEFFNKMKKWEDMANLPPHFRERTERLERNFTVSAVIFKKYEPIFQDIFKYPQEEQPRQQRGRKQRRQPCTTSEIFHFCWVLFIYAKGNFPMISDDLVNSYHLLLCALDLVYGNALQCSNRKELVNPNFKGLSEDCHPKDSKASSDPPCVIEKLCSLHDGLVLEAKGIKEHFWKPYIRKLFEKKLLKGKEENLTGFLEPGNFGESFKAVNKAYEEYVLAAGNLDERVFLGEDAEEEVGTLSRCLSAASGTESAERTQMRDILQQHLDKSKALRVCTPLTGVRYVQENSPCVTPVSTAAHSLSRLHTMLSGLRNAPSEKLERILRSCSRDPTQAIADRLKEMYEIYSQHFQPDENFSNCAKEIANKHFRFAEMLYYKVLESVIEQEQKRLGDMDLSGVLEHDAFHRSLLACCLEVVAFSHKPPGNFPFIAEIFDVPHYHFYKVIEVFIRAEDGLCREVVKHLNQIEEQILDHLAWKTKSPLWDRIRDNENRVPTCEEVMPPQNLERTDEIYIAGSPLTPRRVGEVRADAGGLGRSITSPTTLYDRYSSPTVSTTRRRLFENDSPSEGSTSGRIPPQPLVNAVPVQNVPGETVSVTPVPGQTLVTMATATVTANNGQTVTIPVQGIANENGGITFFPVQVNVGGQAQAVAGSIQPLSAQALAGSLSSQQVTGTTLQVPGPVAIQQISPGGQQQNPGQPLTSSSIRPRKTSSLALFFRKVYYLAGVRLRDLCIKLDISDELRKKIWTCFEFSIIQCTELMMDRHLDQLLMCAIYVMAKVTKEDRSFQNIMRCYRTQPQARSQVYRSVLIKGKRRNSGSSESRSHQNSPTELNTDRASRDSSPVMRSNSTLPVPQPSSAPPTPTRLTGASSDVEEEERGDLIQFYNNIYRKQIQAFAMKYSQANAQTDTPPLSPYPFVRTGSPRRVQLSQSHPIYISPHNNEAMPSPREKIFYYFSNSPSKRLREINSMIRTGETPTKKRGILLDDGSESPAKRICPENHSALLRRLQDVANDRGSQ rbl1_mouse Retinoblastoma-like protein 1 Rbl1 pRb1 107 kDa retinoblastoma-associated protein protein protein mouse Mus musculus Mouse MFEDEPHAEGAAAVAAAREALQALCQELNLDEGSAAEALDDFTAIRGNYSLEGEVIHWLACSLYVACRKSIIPTVGKGVMEGNCVSLTRILRSAKLSLIQFFSKMKKWMDMSNLPQEFRERIERLERNFEVSTVIFKKFEPIFLDIFQNPYEEPPKLPRSRKQRRIPCSVKDLFNFCWTLFVYTKGNFRMIGDDLVNSYHLLLCCLDLIFANAIMCPNRRDLLNPSFKGLPSDFHAPDFKAAEEPPCIIAVLCDLHDGLLVEAKGIKEHYFKPYISKLFDKKILKGECLLDLSSFTDNSKAVNKEYEEYVLTVGDFDERIFLGADAEEEIGTPRKFTADTPFGKLTSQASVECNLQQHFEKKRSFAPSTPLTGRRYLQEKEAVTTPVASATQSVSRLQSIVAGLKSAPSEQLLNIFESCMRNPMGNIIKIVKGIGETFCQHYTQSTDKQPGSHIDFAVNRLKLAEILYYKILETIMVQETRRLHGMDMSVLLEQDIFHKSLMACCLEIVLFAYSSPRTFPWIIEVLDLQPFYFYKVIEVVIRSEEGLSRDMVKHLNSIEEQILESLAWTNNSALWEALHASANRVPSCEEVIFPNNFEIGNGGNVQGHLPMMPMSPIIHPRVKEVRTDSGSLRQDMQPLSPISVHERYSSPAAGSAKRRLFGDDPPKDTLMDKIMAEGTKLKIAPSSVTAESLSISPGQALLTMATTTVTGTTGRKVTVPLHGIANDAGEITLVPISMNPTQESTAESPVSLTAQSLIGTSPKQTHLTKAQDAHLTGVSKPKRTGSLALFYRKVYHLASVRLRDLCLKLDVSNELRRKIWTCFEFTLVHCPDLMKDRHLDQLLLCAFYIMAKVTKEERTFQEIMKSYRNQPQANSHVYRSVLLKSIPGGVVVYNGDCEMTDGDIEDATKTPNCSSEPVKEERGDLIKFYNTVYVGRVKSFALKYDLSNQDHIMDAPPLSPFPHIKQQPGSPRRISQQHSLYVSPHKNGAGLTPRSALLYKFNGSPSKSLKDINNMIRQGEQKTKKRVIAISGDADSPAKRLCQENDDVLLKRLQDVVSERANH rb_human Retinoblastoma-associated protein pRb pp110 p105-Rb RB1 p110-RB1 protein protein human Homo sapiens Human MPPKTPRKTAATAAAAAAEPPAPPPPPPPEEDPEQDSGPEDLPLVRLEFEETEEPDFTALCQKLKIPDHVRERAWLTWEKVSSVDGVLGGYIQKKKELWGICIFIAAVDLDEMSFTFTELQKNIEISVHKFFNLLKEIDTSTKVDNAMSRLLKKYDVLFALFSKLERTCELIYLTQPSSSISTEINSALVLKVSWITFLLAKGEVLQMEDDLVISFQLMLCVLDYFIKLSPPMLLKEPYKTAVIPINGSPRTPRRGQNRSARIAKQLENDTRIIEVLCKEHECNIDEVKNVYFKNFIPFMNSLGLVTSNGLPEVENLSKRYEEIYLKNKDLDARLFLDHDKTLQTDSIDSFETQRTPRKSNLDEEVNVIPPHTPVRTVMNTIQQLMMILNSASDQPSENLISYFNNCTVNPKESILKRVKDIGYIFKEKFAKAVGQGCVEIGSQRYKLGVRLYYRVMESMLKSEEERLSIQNFSKLLNDNIFHMSLLACALEVVMATYSRSTSQNLDSGTDLSFPWILNVLNLKAFDFYKVIESFIKAEGNLTREMIKHLERCEHRIMESLAWLSDSPLFDLIKQSKDREGPTDHLESACPLNLPLQNNHTAADMYLSPVRSPKKKGSTTRVNSTANAETQATSAFQTQKPLKSTSLSLFYKKVYRLAYLRLNTLCERLLSEHPELEHIIWTLFQHTLQNEYELMRDRHLDQIMMCSMYGICKVKNIDLKFKIIVTAYKDLPHAVQETFKRVLIKEEEYDSIIVFYNSVFMQRLKTNILQYASTRPPTLSPIPHIPRSPYKFPSSPLRIPGGNIYISPLKSPYKISEGLPTPTKMTPRSRILVSIGESFGTSEKFQKINQMVCNSDRVLKRSAEGSNPPKPLKKLRFDIEGSDEADGSKHLPGESKFQQKLAEMTSTRTRMQKQKMNDSMDTSNKEEK rbl1_human Retinoblastoma-like protein 1 RBL1 pRb1 107 kDa retinoblastoma-associated protein protein protein human Homo sapiens Human MFEDKPHAEGAAVVAAAGEALQALCQELNLDEGSAAEALDDFTAIRGNYSLEGEVTHWLACSLYVACRKSIIPTVGKGIMEGNCVSLTRILRSAKLSLIQFFSKMKKWMDMSNLPQEFRERIERLERNFEVSTVIFKKYEPIFLDIFQNPYEEPPKLPRSRKQRRIPCSVKDLFNFCWTLFVYTKGNFRMIGDDLVNSYHLLLCCLDLIFANAIMCPNRQDLLNPSFKGLPSDFHTADFTASEEPPCIIAVLCELHDGLLVEAKGIKEHYFKPYISKLFDRKILKGECLLDLSSFTDNSKAVNKEYEEYVLTVGDFDERIFLGADAEEEIGTPRKFTRDTPLGKLTAQANVEYNLQQHFEKKRSFAPSTPLTGRRYLREKEAVITPVASATQSVSRLQSIVAGLKNAPSDQLINIFESCVRNPVENIMKILKGIGETFCQHYTQSTDEQPGSHIDFAVNRLKLAEILYYKILETVMVQETRRLHGMDMSVLLEQDIFHRSLMACCLEIVLFAYSSPRTFPWIIEVLNLQPFYFYKVIEVVIRSEEGLSRDMVKHLNSIEEQILESLAWSHDSALWEALQVSANKVPTCEEVIFPNNFETGNGGNVQGHLPLMPMSPLMHPRVKEVRTDSGSLRRDMQPLSPISVHERYSSPTAGSAKRRLFGEDPPKEMLMDKIITEGTKLKIAPSSSITAENVSILPGQTLLTMATAPVTGTTGHKVTIPLHGVANDAGEITLIPLSMNTNQESKVKSPVSLTAHSLIGASPKQTNLTKAQEVHSTGINRPKRTGSLALFYRKVYHLASVRLRDLCLKLDVSNELRRKIWTCFEFTLVHCPDLMKDRHLDQLLLCAFYIMAKVTKEERTFQEIMKSYRNQPQANSHVYRSVLLKSIPREVVAYNKNINDDFEMIDCDLEDATKTPDCSSGPVKEERGDLIKFYNTIYVGRVKSFALKYDLANQDHMMDAPPLSPFPHIKQQPGSPRRISQQHSIYISPHKNGSGLTPRSALLYKFNGSPSKSLKDINNMIRQGEQRTKKRVIAIDSDAESPAKRVCQENDDVLLKRLQDVVSERANH cenpf-rb1-6 CoIP showing in-vivo interaction between cenpf and rb1 800709 n/a 800713 unspecified role unspecified role bait bait n/a 800714 unspecified role unspecified role prey prey physical association physical association false false false Fig 6A. LEK1 forms complexes with all three members of the Rb family in vivo. cenpf-rbl2-1 CoIP showing in-vivo interaction between cenpf and rbl2 800709 n/a 800713 unspecified role unspecified role bait bait n/a 800715 unspecified role unspecified role prey prey physical association physical association false false false Fig 6A. LEK1 forms complexes with all three members of the Rb family in vivo. cenpf-rbl1-2 CoIP showing in-vivo interaction between cenpf and rbl1 800709 n/a 800716 unspecified role unspecified role prey prey n/a 800713 unspecified role unspecified role bait bait physical association physical association false false false Fig 6A. LEK1 forms complexes with all three members of the Rb family in vivo. cenpf-rb1-7 CoIP showing in-vivo interaction between cenpf and rb1 800709 n/a 800713 unspecified role unspecified role prey prey n/a 800714 unspecified role unspecified role bait bait physical association physical association false false false Fig 6B. LEK1 forms complexes with all three members of the Rb family in vivo. cenpf-rbl1-3 CoIP showing in-vivo interaction between cenpf and rbl1 800709 n/a 800713 unspecified role unspecified role prey prey n/a 800716 unspecified role unspecified role bait bait physical association physical association false false false Fig 6B. LEK1 forms complexes with all three members of the Rb family in vivo. cenpf-rbl2-2 CoIP showing in-vivo interaction between cenpf and rbl2 800709 n/a 800713 unspecified role unspecified role bait bait n/a 800715 unspecified role unspecified role prey prey physical association physical association false false false Fig 6B. LEK1 forms complexes with all three members of the Rb family in vivo. cenpf-rb1-2 pull-down showing interaction between cenpf/lek1 and the pocket domains of rb1 800710 n/a 800717 unspecified role unspecified role prey prey c-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 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 false n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 5A and 5B. The C-terminal portion of LEK1 interacts with sequences in all three pocket subdomains of Rb. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rb1-3 pull-down showing interaction between cenpf/lek1 and the pocket domains of rb1 800710 n/a 800717 unspecified role unspecified role prey prey 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 false c-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 5A and 5B. The C-terminal portion of LEK1 interacts with sequences in all three pocket subdomains of Rb. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rb1-4 pull-down showing interaction between cenpf/lek1 and the pocket domains of rb1 800710 n/a 800717 unspecified role unspecified role prey prey 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 false c-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 5A and 5B. The C-terminal portion of LEK1 interacts with sequences in all three pocket subdomains of Rb. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rb1-5 pull-down showing interaction between cenpf/lek1 and the pocket domains of rb1 800710 n/a 800713 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 800717 unspecified role unspecified role prey prey cys-709-phe binding region binding-associated region binding region mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false 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 false c-terminus sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig 5C. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rbl1-1 pull-down showing interaction between cenpf/lek1 and the pocket domains of rbl1 800710 n/a 800718 unspecified role unspecified role prey prey n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 5B. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rb1-1 CoIP showing interaction between cenpf and rb1 800711 n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 800714 unspecified role unspecified role prey prey physical association physical association false false false Fig 3A and 3B. LEK1 associates with not just one, but all three members of the Rb family, suggesting that LEK1 has the potential to affect the activity of all members of the Rb family. The authors have not given the exact positions of the c-terminus they have used in their interaction studies. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rbl1 CoIP showing interaction between cenpf and rbl1 800711 n/a 800716 unspecified role unspecified role prey prey n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 3A and 3B. LEK1 associates with not just one, but all three members of the Rb family, suggesting that LEK1 has the potential to affect the activity of all members of the Rb family. The authors have not given the exact positions of the c-terminus they have used in their interaction studies. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rbl2 CoIP showing interaction between cenpf and rbl2 800711 n/a 800715 unspecified role unspecified role prey prey n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 3A and 3B. LEK1 associates with not just one, but all three members of the Rb family, suggesting that LEK1 has the potential to affect the activity of all members of the Rb family. The authors have not given the exact positions of the c-terminus they have used in their interaction studies. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. cenpf-rb1 pull-down showing interaction between cenpf and rb1 800712 n/a 800714 unspecified role unspecified role prey prey spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection n/a 800713 unspecified role unspecified role bait bait c-terminus required to bind necessary binding region required to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig 2. The C-terminal portion of LEK1 interacts with hypophosphorylated full-length Rb1 The authors have not given the exact positions of the c-terminus they have used in their interaction studies. I have assumed that cenpf used for this interaction is from mouse. The origin of the cenpf protein is deduced from a section in the introduction "The human proteins, Mitosin and CENP-F, associate with the kinetochore complex during mitosis and display an expression pattern that is dependent on cell cycle progression and independent of developmental stage (11, 12). In contrast, the chicken and mouse LEK proteins, CMF1 and LEK1, respectively, display expression patterns that are regulated during development (13, 14)." Also a section of the sequence for each cenpf from the different species is presented in Figure 1. The authors do not state the exact insect cells used in the generation of wt Rb using the Baculoviral expression system but doing a google and pubmed search, the most common insects cells for this are Sf9 cells and so that is what I have stated that the Rb protein is expressed in.