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/ The BRC repeats of BRCA2 modulate the DNA-binding selectivity of RAD51. The BRC repeats of BRCA2 modulate the DNA-binding selectivity of RAD51. Cell (0092-8674) 2009 imex curation Carreira A., Hilario J., Amitani I., Baskin RJ., Shivji MK., Venkitaraman AR., Kowalczykowski SC. Cancer - Interactions investigated in the context of cancer sckowalczykowski@ucdavis.edu 01-Sep-2009: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Carreira A., Hilario J., Amitani I., Baskin RJ., Shivji MK., Venkitaraman AR., Kowalczykowski SC. Cancer - Interactions investigated in the context of cancer sckowalczykowski@ucdavis.edu Accepted 2009-JUL-23 by DAVET. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions imex curation The BRC repeats of BRCA2 modulate the DNA-binding selectivity of RAD51. The BRC repeats of BRCA2 modulate the DNA-binding selectivity of RAD51. Cell (0092-8674) 2009 imex curation Carreira A., Hilario J., Amitani I., Baskin RJ., Shivji MK., Venkitaraman AR., Kowalczykowski SC. Cancer - Interactions investigated in the context of cancer sckowalczykowski@ucdavis.edu 01-Sep-2009: Contacted by JYOTI. davet Only protein-protein interactions in vitro In vitro atpase assay atpase assay predetermined predetermined participant predetermined Carreira A., Hilario J., Amitani I., Baskin RJ., Shivji MK., Venkitaraman AR., Kowalczykowski SC. Cancer - Interactions investigated in the context of cancer sckowalczykowski@ucdavis.edu Accepted 2009-JUL-23 by DAVET. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions imex curation dt40_dna 40-mer poly-dT (dT40) ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT 4B1B1B5F5F5F5F5F dt90_dna 90-mer poly-dT (dT90) ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT 0F5F0E1A4B0E1A4B phix174_dsdna Enterobacteria phage phiX174 linear dsDNA ds dna double stranded deoxyribonucleic acid ds DNA ds dna bpphx Enterobacteria phage phiX174 Bacteriophage phi-X174 phix174_ssdna Enterobacteria phage phiX174 genome ss dna single stranded deoxyribonucleic acid ss DNA ss dna bpphx Enterobacteria phage phiX174 Bacteriophage phi-X174 atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) m13 Phage M13 genome dna deoxyribonucleic acid DNA deoxyribonucleic acid dna DNA deoxyribonucleic acid bpm13 Enterobacteria phage M13 Bacteriophage M13 rad51_human DNA repair protein RAD51 homolog 1 RAD51 RAD51A RECA RAD51 homolog A protein protein human Homo sapiens Human MAMQMQLEANADTSVEEESFGPQPISRLEQCGINANDVKKLEEAGFHTVEAVAYAPKKELINIKGISEAKADKILAEAAKLVPMGFTTATEFHQRRSEIIQITTGSKELDKLLQGGIETGSITEMFGEFRTGKTQICHTLAVTCQLPIDRGGGEGKAMYIDTEGTFRPERLLAVAERYGLSGSDVLDNVAYARAFNTDHQTQLLYQASAMMVESRYALLIVDSATALYRTDYSGRGELSARQMHLARFLRMLLRLADEFGVAVVITNQVVAQVDGAAMFAADPKKPIGGNIIAHASTTRLYLRKGRGETRICKIYDSPCLPEAEAMFAINADGVGDAKD DIP protein Q06609 original sequence version: 154 DIP protein Q06609 original sequence version: 145 DIP protein Q06609 original sequence version: 122 DIP protein Q06609 original sequence version: 121 DIP protein Q06609 original sequence version: 125 DIP protein Q06609 original sequence version: 123 DIP protein Q06609 original sequence version: 144 DIP protein Q06609 original sequence version: 167 DIP protein Q06609 original sequence version: 169 26578E6206DEDEDA brca2_human Breast cancer type 2 susceptibility protein BRCA2 FANCD1 FACD Fanconi anemia group D1 protein protein protein human Homo sapiens Human MPIGSKERPTFFEIFKTRCNKADLGPISLNWFEELSSEAPPYNSEPAEESEHKNNNYEPNLFKTPQRKPSYNQLASTPIIFKEQGLTLPLYQSPVKELDKFKLDLGRNVPNSRHKSLRTVKTKMDQADDVSCPLLNSCLSESPVVLQCTHVTPQRDKSVVCGSLFHTPKFVKGRQTPKHISESLGAEVDPDMSWSSSLATPPTLSSTVLIVRNEEASETVFPHDTTANVKSYFSNHDESLKKNDRFIASVTDSENTNQREAASHGFGKTSGNSFKVNSCKDHIGKSMPNVLEDEVYETVVDTSEEDSFSLCFSKCRTKNLQKVRTSKTRKKIFHEANADECEKSKNQVKEKYSFVSEVEPNDTDPLDSNVANQKPFESGSDKISKEVVPSLACEWSQLTLSGLNGAQMEKIPLLHISSCDQNISEKDLLDTENKRKKDFLTSENSLPRISSLPKSEKPLNEETVVNKRDEEQHLESHTDCILAVKQAISGTSPVASSFQGIKKSIFRIRESPKETFNASFSGHMTDPNFKKETEASESGLEIHTVCSQKEDSLCPNLIDNGSWPATTTQNSVALKNAGLISTLKKKTNKFIYAIHDETSYKGKKIPKDQKSELINCSAQFEANAFEAPLTFANADSGLLHSSVKRSCSQNDSEEPTLSLTSSFGTILRKCSRNETCSNNTVISQDLDYKEAKCNKEKLQLFITPEADSLSCLQEGQCENDPKSKKVSDIKEEVLAAACHPVQHSKVEYSDTDFQSQKSLLYDHENASTLILTPTSKDVLSNLVMISRGKESYKMSDKLKGNNYESDVELTKNIPMEKNQDVCALNENYKNVELLPPEKYMRVASPSRKVQFNQNTNLRVIQKNQEETTSISKITVNPDSEELFSDNENNFVFQVANERNNLALGNTKELHETDLTCVNEPIFKNSTMVLYGDTGDKQATQVSIKKDLVYVLAEENKNSVKQHIKMTLGQDLKSDISLNIDKIPEKNNDYMNKWAGLLGPISNHSFGGSFRTASNKEIKLSEHNIKKSKMFFKDIEEQYPTSLACVEIVNTLALDNQKKLSKPQSINTVSAHLQSSVVVSDCKNSHITPQMLFSKQDFNSNHNLTPSQKAEITELSTILEESGSQFEFTQFRKPSYILQKSTFEVPENQMTILKTTSEECRDADLHVIMNAPSIGQVDSSKQFEGTVEIKRKFAGLLKNDCNKSASGYLTDENEVGFRGFYSAHGTKLNVSTEALQKAVKLFSDIENISEETSAEVHPISLSSSKCHDSVVSMFKIENHNDKTVSEKNNKCQLILQNNIEMTTGTFVEEITENYKRNTENEDNKYTAASRNSHNLEFDGSDSSKNDTVCIHKDETDLLFTDQHNICLKLSGQFMKEGNTQIKEDLSDLTFLEVAKAQEACHGNTSNKEQLTATKTEQNIKDFETSDTFFQTASGKNISVAKESFNKIVNFFDQKPEELHNFSLNSELHSDIRKNKMDILSYEETDIVKHKILKESVPVGTGNQLVTFQGQPERDEKIKEPTLLGFHTASGKKVKIAKESLDKVKNLFDEKEQGTSEITSFSHQWAKTLKYREACKDLELACETIEITAAPKCKEMQNSLNNDKNLVSIETVVPPKLLSDNLCRQTENLKTSKSIFLKVKVHENVEKETAKSPATCYTNQSPYSVIENSALAFYTSCSRKTSVSQTSLLEAKKWLREGIFDGQPERINTADYVGNYLYENNSNSTIAENDKNHLSEKQDTYLSNSSMSNSYSYHSDEVYNDSGYLSKNKLDSGIEPVLKNVEDQKNTSFSKVISNVKDANAYPQTVNEDICVEELVTSSSPCKNKNAAIKLSISNSNNFEVGPPAFRIASGKIVCVSHETIKKVKDIFTDSFSKVIKENNENKSKICQTKIMAGCYEALDDSEDILHNSLDNDECSTHSHKVFADIQSEEILQHNQNMSGLEKVSKISPCDVSLETSDICKCSIGKLHKSVSSANTCGIFSTASGKSVQVSDASLQNARQVFSEIEDSTKQVFSKVLFKSNEHSDQLTREENTAIRTPEHLISQKGFSYNVVNSSAFSGFSTASGKQVSILESSLHKVKGVLEEFDLIRTEHSLHYSPTSRQNVSKILPRVDKRNPEHCVNSEMEKTCSKEFKLSNNLNVEGGSSENNHSIKVSPYLSQFQQDKQQLVLGTKVSLVENIHVLGKEQASPKNVKMEIGKTETFSDVPVKTNIEVCSTYSKDSENYFETEAVEIAKAFMEDDELTDSKLPSHATHSLFTCPENEEMVLSNSRIGKRRGEPLILVGEPSIKRNLLNEFDRIIENQEKSLKASKSTPDGTIKDRRLFMHHVSLEPITCVPFRTTKERQEIQNPNFTAPGQEFLSKSHLYEHLTLEKSSSNLAVSGHPFYQVSATRNEKMRHLITTGRPTKVFVPPFKTKSHFHRVEQCVRNINLEENRQKQNIDGHGSDDSKNKINDNEIHQFNKNNSNQAAAVTFTKCEEEPLDLITSLQNARDIQDMRIKKKQRQRVFPQPGSLYLAKTSTLPRISLKAAVGGQVPSACSHKQLYTYGVSKHCIKINSKNAESFQFHTEDYFGKESLWTGKGIQLADGGWLIPSNDGKAGKEEFYRALCDTPGVDPKLISRIWVYNHYRWIIWKLAAMECAFPKEFANRCLSPERVLLQLKYRYDTEIDRSRRSAIKKIMERDDTAAKTLVLCVSDIISLSANISETSSNKTSSADTQKVAIIELTDGWYAVKAQLDPPLLAVLKNGRLTVGQKIILHGAELVGSPDACTPLEAPESLMLKISANSTRPARWYTKLGFFPDPRPFPLPLSSLFSDGGNVGCVDVIIQRAYPIQWMEKTSSGLYIFRNEREEEKEAAKYVEAQQKRLEALFTKIQEEFEEHEENTTKPYLPSRALTRQQVRALQDGAELYEAVKNAADPAYLEGYFSEEQLRALNNHRQMLNDKKQAQIQLEIRKAMESAEQKEQGLSRDVTTVWKLRIVSYSKKEKDSVILSIWRPSSDLYSLLTEGKRYRIYHLATSKSKSKSERANIQLAATKKTQYQQLPVSDEILFQIYQPREPLHFSKFLDPDFQPSCSEVDLIGFVVSVVKKTGLAPFVYLSDECYNLLAIKFWIDLNEDIIKPHMLIAASNLQWRPESKSGLLTLFAGDFSVFSASPKEGHFQETFNKMKNTVENIDILCNEAENKLMHILHANDPKWSTPTKDCTSGPYTAQIIPGTGNKLLMSSPNCEIYYQSPLSLCMAKRKSVSTPVSAQMTSKSCKGEKEIDDQKNCKKRRALDFLSRLPLPPPVSPICTFVSPAAQKAFQPPRSCGTKYETPIKKKELNSPQMTPFKKFNEISLLESNSIADEELALINTQALLSGSTGEKQFISVSESTRTAPTSSEDYLRLKRRCTTSLIKEQESSQASTEECEKNKQDTITTKKYI DIP protein P51587 original sequence version: 162 DIP protein P51587 original sequence version: 83 DIP protein P51587 original sequence version: 145 DIP protein P51587 original sequence version: 122 DIP protein P51587 original sequence version: 120 DIP protein P51587 original sequence version: 138 DIP protein P51587 original sequence version: 125 DIP protein P51587 original sequence version: 123 DIP protein P51587 original sequence version: 155 DIP protein P51587 original sequence version: 168 DIP protein P51587 original sequence version: 156 DIP protein P51587 original sequence version: 180 6A0B3B7B332153EB dt60_dna 60-mer poly-dT (dT60) ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT 0F5F5F5F0B5F5F0B rad51-dt40-1 1 3 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction In the presence of ATP, the RAD51 nucleoprotein filaments are partially stable and only not, vert, similar30% of the DNA is shifted (Figure 1A, lane 2). Fig. 1A dT40 is 32P labelled. rad51-dt40-2 1 10 stimulator stimulator ancillary ancillary P51587:p.Thr1526Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T A Wed May 02 18:45:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr1526ala' Tue Jun 30 20:08:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr1526Ala' 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 brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role prey prey physical association physical association The addition of BRC4 unexpectedly produced a BRC4 concentration-dependent increase in RAD51-ssDNA complexes from 30% to almost 100% (lanes 3�8), Fig. 1 dT40 is 32P labelled. Interaction required Mg2+ and ATP. In the absence of ATP, BRCA2 inhibited filament formation. rad51-atp-1 2 7 enzyme target enzyme target substrate substrate neutral component neutral component 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 3 unspecified role unspecified role ancillary ancillary atpase reaction atpase reaction ssDNA-dependent ATPase activity of RAD51 Fig. 2A ATP is 32P labelled. rad51-atp-2 2 7 enzyme target enzyme target substrate substrate neutral component neutral component 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role ancillary ancillary atpase reaction atpase reaction ssDNA-dependent ATPase activity of RAD51 Fig. 2A ATP is 32P labelled. rad51-atp-3 2 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 4 unspecified role unspecified role ancillary ancillary atpase reaction atpase reaction ssDNA-dependent ATPase activity of RAD51 Fig. 2A ATP is 32P labelled. rad51-atp-4 2 10 inhibitor inhibitor neutral component neutral component brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis 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 ancillary ancillary 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component atpase reaction atpase reaction BRC4 reduced ATP hydrolysis by RAD51 for all the ssDNA examined, both homopolymers and natural DNA (Figure 2A). Fig. 2A ATP is 32P labelled. rad51-atp-5 2 7 enzyme target enzyme target substrate substrate neutral component neutral component 10 inhibitor inhibitor neutral component neutral component 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 brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 11 unspecified role unspecified role ancillary ancillary atpase reaction atpase reaction BRC4 reduced ATP hydrolysis by RAD51 for all the ssDNA examined, both homopolymers and natural DNA (Figure 2A). Fig. 2A ATP is 32P labelled. rad51-atp-6 2 10 inhibitor inhibitor neutral component neutral component brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role ancillary ancillary 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component atpase reaction atpase reaction BRC4 reduced ATP hydrolysis by RAD51 for all the ssDNA examined, both homopolymers and natural DNA (Figure 2A). Fig. 2A ATP is 32P labelled. rad51-atp-7 2 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 10 inhibitor inhibitor neutral component neutral component 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 brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role ancillary ancillary atpase reaction atpase reaction BRC4 reduced ATP hydrolysis by RAD51 for all the ssDNA examined, both homopolymers and natural DNA (Figure 2A). Fig. 2A ATP is 32P labelled. rad51-atp-8 2 8 unspecified role unspecified role ancillary ancillary 7 enzyme target enzyme target substrate substrate neutral component neutral component 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) atpase reaction atpase reaction ssDNA-dependent ATPase activity of RAD51 Fig. 2A ATP is 32P labelled. rad51-atp-9 2 6 unspecified role unspecified role ancillary ancillary 10 inhibitor inhibitor neutral component neutral component brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis 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) 7 enzyme target enzyme target substrate substrate neutral component neutral component 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) atpase reaction atpase reaction BRC4 reduced ATP hydrolysis by RAD51 for all the ssDNA examined, both homopolymers and natural DNA (Figure 2A). Fig. 2A ATP is 32P labelled. rad51-atp-10 2 6 unspecified role unspecified role ancillary ancillary 9 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component atpase reaction atpase reaction ssDNA-dependent ATPase activity of RAD51 Fig. 2A ATP is 32P labelled. rad51-phix174-1 1 9 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait direct interaction direct interaction binding of RAD51 to dsDNA. Fig. 3A phiX174 is 32P labelled. rad51-phix174-2 1 9 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 10 inhibitor inhibitor ancillary ancillary P51587:p.Thr1526Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T A Wed May 02 16:19:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr1526ala' Tue Jun 30 17:29:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr1526Ala' 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 brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey physical association physical association However, in the presence of ATP, increasing concentrations of BRC4 inhibited binding of RAD51 to dsDNA, which is opposite to the behavior seen with ssDNA. Figs. 3A, 3B, Supp. Fig. 4B phiX174 is 32P labelled. Interaction required ATP. rad51-dt40-3 1 3 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 10 stimulator stimulator ancillary ancillary brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Untagged BRC4 stimulates binding of RAD51 to ssDNA Supp. Fig. 2 dT40 is 32P labelled. rad51-phix174-3 1 5 unspecified role unspecified role prey prey 10 inhibitor inhibitor ancillary ancillary brc4 repeat region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) physical association physical association Both tagged and untagged BRC4 inhibit binding of RAD51 to dsDNA. Supp. Fig. 4 phiX174 is 32P labelled. Interaction required ATP.