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/ Interactions of human hMSH2 with hMSH3 and hMSH2 with hMSH6: examination of mutations found in hereditary nonpolyposis colorectal cancer. Interactions of human hMSH2 with hMSH3 and hMSH2 with hMSH6: examination of mutations found in hereditary nonpolyposis colorectal cancer. Mol. Cell. Biol. (0270-7306) 1998 Guerrette S., Wilson T., Gradia S., Fishel R. Cancer - Interactions investigated in the context of cancer rfishel@hendrix.jci.tju.edu 27-NOV-2006: Contacted by JYOTI. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Guerrette S., Wilson T., Gradia S., Fishel R. Cancer - Interactions investigated in the context of cancer rfishel@hendrix.jci.tju.edu Accepted 2006-NOV-13 by DAVET. 27-NOV-2006: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 1998 msh3_human DNA mismatch repair protein Msh3 Divergent upstream protein Mismatch repair protein 1 MSH3 DUC1 DUG protein protein human Homo sapiens Human MSRRKPASGGLAASSSAPARQAVLSRFFQSTGSLKSTSSSTGAADQVDPGAAAAAAAAAAAAPPAPPAPAFPPQLPPHIATEIDRRKKRPLENDGPVKKKVKKVQQKEGGSDLGMSGNSEPKKCLRTRNVSKSLEKLKEFCCDSALPQSRVQTESLQERFAVLPKCTDFDDISLLHAKNAVSSEDSKRQINQKDTTLFDLSQFGSSNTSHENLQKTASKSANKRSKSIYTPLELQYIEMKQQHKDAVLCVECGYKYRFFGEDAEIAARELNIYCHLDHNFMTASIPTHRLFVHVRRLVAKGYKVGVVKQTETAALKAIGDNRSSLFSRKLTALYTKSTLIGEDVNPLIKLDDAVNVDEIMTDTSTSYLLCISENKENVRDKKKGNIFIGIVGVQPATGEVVFDSFQDSASRSELETRMSSLQPVELLLPSALSEQTEALIHRATSVSVQDDRIRVERMDNIYFEYSHAFQAVTEFYAKDTVDIKGSQIISGIVNLEKPVICSLAAIIKYLKEFNLEKMLSKPENFKQLSSKMEFMTINGTTLRNLEILQNQTDMKTKGSLLWVLDHTKTSFGRRKLKKWVTQPLLKLREINARLDAVSEVLHSESSVFGQIENHLRKLPDIERGLCSIYHKKCSTQEFFLIVKTLYHLKSEFQAIIPAVNSHIQSDLLRTVILEIPELLSPVEHYLKILNEQAAKVGDKTELFKDLSDFPLIKKRKDEIQGVIDEIRMHLQEIRKILKNPSAQYVTVSGQEFMIEIKNSAVSCIPTDWVKVGSTKAVSRFHSPFIVENYRHLNQLREQLVLDCSAEWLDFLEKFSEHYHSLCKAVHHLATVDCIFSLAKVAKQGDYCRPTVQEERKIVIKNGRHPVIDVLLGEQDQYVPNNTDLSEDSERVMIITGPNMGGKSSYIKQVALITIMAQIGSYVPAEEATIGIVDGIFTRMGAADNIYKGQSTFMEELTDTAEIIRKATSQSLVILDELGRGTSTHDGIAIAYATLEYFIRDVKSLTLFVTHYPPVCELEKNYSHQVGNYHMGFLVSEDESKLDPGAAEQVPDFVTFLYQITRGIAARSYGLNVAKLADVPGEILKKAAHKSKELEGLINTKRKRLKYFAKLWTMHNAQDLQKWTEEFNMEETQTSLLH DIP protein P20585 original sequence version: 125 DIP protein P20585 original sequence version: 97 DIP protein P20585 original sequence version: 111 FBAE6B84D3F86032 msh2_human DNA mismatch repair protein Msh2 MSH2 MutS protein homolog 2 protein protein human Homo sapiens Human MAVQPKETLQLESAAEVGFVRFFQGMPEKPTTTVRLFDRGDFYTAHGEDALLAAREVFKTQGVIKYMGPAGAKNLQSVVLSKMNFESFVKDLLLVRQYRVEVYKNRAGNKASKENDWYLAYKASPGNLSQFEDILFGNNDMSASIGVVGVKMSAVDGQRQVGVGYVDSIQRKLGLCEFPDNDQFSNLEALLIQIGPKECVLPGGETAGDMGKLRQIIQRGGILITERKKADFSTKDIYQDLNRLLKGKKGEQMNSAVLPEMENQVAVSSLSAVIKFLELLSDDSNFGQFELTTFDFSQYMKLDIAAVRALNLFQGSVEDTTGSQSLAALLNKCKTPQGQRLVNQWIKQPLMDKNRIEERLNLVEAFVEDAELRQTLQEDLLRRFPDLNRLAKKFQRQAANLQDCYRLYQGINQLPNVIQALEKHEGKHQKLLLAVFVTPLTDLRSDFSKFQEMIETTLDMDQVENHEFLVKPSFDPNLSELREIMNDLEKKMQSTLISAARDLGLDPGKQIKLDSSAQFGYYFRVTCKEEKVLRNNKNFSTVDIQKNGVKFTNSKLTSLNEEYTKNKTEYEEAQDAIVKEIVNISSGYVEPMQTLNDVLAQLDAVVSFAHVSNGAPVPYVRPAILEKGQGRIILKASRHACVEVQDEIAFIPNDVYFEKDKQMFHIITGPNMGGKSTYIRQTGVIVLMAQIGCFVPCESAEVSIVDCILARVGAGDSQLKGVSTFMAEMLETASILRSATKDSLIIIDELGRGTSTYDGFGLAWAISEYIATKIGAFCMFATHFHELTALANQIPTVNNLHVTALTTEETLTMLYQVKKGVCDQSFGIHVAELANFPKHVIECAKQKALELEEFQYIGESQGYDIMEPAAKKCYLEREQGEKIIQEFLSKVKQMPFTEMSEENITIKLKQLKAEVIAKNNSFVNEIISRIKVTT DIP protein P43246 original sequence version: 156 DIP protein P43246 original sequence version: 123 DIP protein P43246 original sequence version: 138 664A058C78242E05 msh6_human DNA mismatch repair protein Msh6 MSH6 MutS-alpha 160 kDa subunit G/T mismatch-binding protein GTBP MutS protein homolog 6 protein protein human Homo sapiens Human MSRQSTLYSFFPKSPALSDANKASARASREGGRAAAAPGASPSPGGDAAWSEAGPGPRPLARSASPPKAKNLNGGLRRSVAPAAPTSCDFSPGDLVWAKMEGYPWWPCLVYNHPFDGTFIREKGKSVRVHVQFFDDSPTRGWVSKRLLKPYTGSKSKEAQKGGHFYSAKPEILRAMQRADEALNKDKIKRLELAVCDEPSEPEEEEEMEVGTTYVTDKSEEDNEIESEEEVQPKTQGSRRSSRQIKKRRVISDSESDIGGSDVEFKPDTKEEGSSDEISSGVGDSESEGLNSPVKVARKRKRMVTGNGSLKRKSSRKETPSATKQATSISSETKNTLRAFSAPQNSESQAHVSGGGDDSSRPTVWYHETLEWLKEEKRRDEHRRRPDHPDFDASTLYVPEDFLNSCTPGMRKWWQIKSQNFDLVICYKVGKFYELYHMDALIGVSELGLVFMKGNWAHSGFPEIAFGRYSDSLVQKGYKVARVEQTETPEMMEARCRKMAHISKYDRVVRREICRIITKGTQTYSVLEGDPSENYSKYLLSLKEKEEDSSGHTRAYGVCFVDTSLGKFFIGQFSDDRHCSRFRTLVAHYPPVQVLFEKGNLSKETKTILKSSLSCSLQEGLIPGSQFWDASKTLRTLLEEEYFREKLSDGIGVMLPQVLKGMTSESDSIGLTPGEKSELALSALGGCVFYLKKCLIDQELLSMANFEEYIPLDSDTVSTTRSGAIFTKAYQRMVLDAVTLNNLEIFLNGTNGSTEGTLLERVDTCHTPFGKRLLKQWLCAPLCNHYAINDRLDAIEDLMVVPDKISEVVELLKKLPDLERLLSKIHNVGSPLKSQNHPDSRAIMYEETTYSKKKIIDFLSALEGFKVMCKIIGIMEEVADGFKSKILKQVISLQTKNPEGRFPDLTVELNRWDTAFDHEKARKTGLITPKAGFDSDYDQALADIRENEQSLLEYLEKQRNRIGCRTIVYWGIGRNRYQLEIPENFTTRNLPEEYELKSTKKGCKRYWTKTIEKKLANLINAEERRDVSLKDCMRRLFYNFDKNYKDWQSAVECIAVLDVLLCLANYSRGGDGPMCRPVILLPEDTPPFLELKGSRHPCITKTFFGDDFIPNDILIGCEEEEQENGKAYCVLVTGPNMGGKSTLMRQAGLLAVMAQMGCYVPAEVCRLTPIDRVFTRLGASDRIMSGESTFFVELSETASILMHATAHSLVLVDELGRGTATFDGTAIANAVVKELAETIKCRTLFSTHYHSLVEDYSQNVAVRLGHMACMVENECEDPSQETITFLYKFIKGACPKSYGFNAARLANLPEEVIQKGHRKAREFEKMNQSLRLFREVCLASERSTVDAEAVHKLLTLIKEL DIP protein P52701 original sequence version: 128 4A4AA9F8ECB8FFE9 msh2-msh3 1 2 unspecified role unspecified role prey prey region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 3 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 2 The interaction can be mediated by either of two regions in MSH3, one C-terminal and the other N-terminal. The results indicate that the C-terminal region of MSH3 interacts with the C-terminal region of MSH2 and the N-terminal region of MSH3 interacts with the N-terminal region of MSH2. The aminoterminal region was resolved between aa 126 and 250 (Fig. 2, pairs 6 to 9). It is important to note that we found the level of IVTT expression to be insufficient with polypeptides that contained less than 100 aa (data not shown). Thus, in order to fully map the carboxy-terminal region we adopted an internal deletion strategy. Using this strategy, the carboxyl interaction region was localized between aa 1050 and 1128 The HeLa cDNA clone of hMSH3 used here has a translation with an insertion of 9 residues (AAAAAAAPP) between residues 56 and 57 of the UniprotkB entry for hMSH3 (P20585). msh3-msh2-1 1 2 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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) 3 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association 4 The interaction can be mediated by either of two regions in MSH2, one C-terminal and the other N-terminal. The results indicate that the C-terminal region of MSH3 interacts with the C-terminal region of MSH2 and the N-terminal region of MSH3 interacts with the N-terminal region of MSH2. The HeLa cDNA clone of hMSH3 used here has a translation with an insertion of 9 residues (AAAAAAAPP) between residues 56 and 57 of the UniprotkB entry for hMSH3 (P20585). GST-hMSH3 (aa 1 to 297) interacted most strongly with aa 251 to 750 of hMSH2 Note a faint band also appears corresponding to MSH2 aa 1-498. msh2-msh6 1 4 unspecified role unspecified role prey prey region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 3 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 5 The interaction can be mediated by either of two regions in MSH6, one N-terminal and one C-terminal. The results indicate that the C-terminal region of MSH6 interacts with the C-terminal region of MSH2 and the N-terminal region of MSH6 interacts with the N-terminal region of MSH2. hMSH6 contained two interaction regions with hMSH2 (Fig. 5, pairs 1 to 6). The amino-terminal interaction region was mapped from aa 326 to 575 (Fig. 5, pairs 7 to 10). The carboxy-terminal interaction region lies between aa 953 and 1360 msh6-msh2 1 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association 7 The results indicate that the C-terminal region of MSH6 interacts with the C-terminal region of MSH2 and the N-terminal region of MSH6 interacts with the N-terminal region of MSH2. The results suggest the same regions of MSH2 are involved in the interaction with MSH3 and MSH6. There is also a faint band corresponding to hMSH2 aa 1-498. msh2-msh2 1 3 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 c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role prey prey region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association Using a GST-hMSH2 (aa 751 to 934) construct we found that this C-terminal region interacted with the carboxy terminus of hMSH2 (data not shown). This construct displayed specific interaction with the hMSH2 truncation mutants aa 751 to 934 and aa 700 to 934 (D800 to 875). However, it did not interact with either the hMSH2 truncation mutant aa 751 to 900 or aa 700 to 934 (D800 to 900). Thus, the hMSH2 homodimer appears to display the same C-terminal interaction pattern that we observed with hMSH2 binding to hMSH3 or hMSH6 (Fig. 4D and 7D, pairs 6 to 10), implicating aa 875 to 934 of hMSH2 in selfassociation The HeLa cDNA clone of hMSH3 used here has a translation with an insertion of 9 residues (AAAAAAAPP) between residues 56 and 57 of the UniprotkB entry for hMSH3 (P20585). msh3-msh2-2 1 3 unspecified role unspecified role prey prey c-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 2 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 4 The results indicate that the C-terminal region of MSH3 interacts with the C-terminal region of MSH2 and the N-terminal region of MSH3 interacts with the N-terminal region of MSH2. The HeLa cDNA clone of hMSH3 used here has a translation with an insertion of 9 residues (AAAAAAAPP) between residues 56 and 57 of the UniprotkB entry for hMSH3 (P20585). The GST-hMSH3 (aa 1025 to 1128) truncation also allowed us to further resolve the carboxy-terminal interaction region of hMSH2 to aa 875 to 934 msh6-msh2-1 1 4 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1164915]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 1660-1660, sequence length 1360 [EBI-1164915]1302-1660 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) 3 unspecified role unspecified role prey prey c-terminal required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro physical association physical association 7 The results indicate that the C-terminal region of MSH6 interacts with the C-terminal region of MSH2 and the N-terminal region of MSH6 interacts with the N-terminal region of MSH2. The results suggest the same regions of MSH2 are involved in the interaction with MSH3 and MSH6. There is also a faint band corresponding to hMSH2 aa 1-498. By using a GST fusion protein which contained a truncation mutant of hMSH6 containing aa 1302 to 1360 (similar to the GST-hMSH3 truncation system described above), the carboxyterminal interaction region of hMSH2 was localized to aa 875 to 934 msh3-msh2-3 1 3 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 35S-methionine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 2 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 8 The HeLa cDNA clone of hMSH3 used here has a translation with an insertion of 9 residues (AAAAAAAPP) between residues 56 and 57 of the UniprotkB entry for hMSH3 (P20585). one of these HNPCC mutant constructs, R524P (aa 499 to 650), appeared to show reduced interaction