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/ mortier_bar-2007-1 A key presynaptic role in transformation for a widespread bacterial protein: DprA conveys incoming ssDNA to RecA. 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 Mortier-Barriere I., Velten M., Dupaigne P., Mirouze N., Pietrement O., McGovern S., Fichant G., Martin B., Noirot P., Le Cam E., Polard P., Claverys JP. patrice.polard@ibcg.biotoul.fr,claverys@ibcg.biotoul.fr Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation mortier_bar-2007-2 A key presynaptic role in transformation for a widespread bacterial protein: DprA conveys incoming ssDNA to RecA. 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 predetermined predetermined participant predetermined Mortier-Barriere I., Velten M., Dupaigne P., Mirouze N., Pietrement O., McGovern S., Fichant G., Martin B., Noirot P., Le Cam E., Polard P., Claverys JP. patrice.polard@ibcg.biotoul.fr,claverys@ibcg.biotoul.fr Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation mortier_bar-2007-3 A key presynaptic role in transformation for a widespread bacterial protein: DprA conveys incoming ssDNA to RecA. strpn Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4) pull down pull down ms participant Identification by mass spectrometry ms participant Mortier-Barriere I., Velten M., Dupaigne P., Mirouze N., Pietrement O., McGovern S., Fichant G., Martin B., Noirot P., Le Cam E., Polard P., Claverys JP. patrice.polard@ibcg.biotoul.fr,claverys@ibcg.biotoul.fr Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation mortier_bar-2007-4 A key presynaptic role in transformation for a widespread bacterial protein: DprA conveys incoming ssDNA to RecA. in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Mortier-Barriere I., Velten M., Dupaigne P., Mirouze N., Pietrement O., McGovern S., Fichant G., Martin B., Noirot P., Le Cam E., Polard P., Claverys JP. patrice.polard@ibcg.biotoul.fr,claverys@ibcg.biotoul.fr Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation a0a0h2uqa6_strpn SP_1266 protein protein strpn Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4) MELFMKITNYEIYKLKKSGLTNQQILKVLEYGENVDQELLLGDIADISGCRNPAVFMERYFQIDDAHLSKEFQKFPSFSILDDCYPWDLSEIYDAPVLLFYKGNLDLLKFPKVAVVGSRACSKQGAKSVEKVIQGLENELVIVSGLAKGIDTAAHMAALQNGGKTIAVIGTGLDVFYPKANKRLQDYIGNDHLVLSEYGPGEQPLKFHFPARNRIIAGLCRGVIVAEAKMRSGSLITCERAMEEGRDVFAIPGSILDGLSDGCHHLIQEGAKLVTSGQDVLAEFEF dt90_dna 90-mer poly-dT (dT90) ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT 80mer_random_ssdna 80-mer oligonucleotide of random sequence ss dna single stranded deoxyribonucleic acid ss DNA ss dna chemical synthesis Chemical synthesis (Chemical synthesis) This is an 80-mer (ssDNA) of random sequence. genome_bpphx_dna Complete genome of bacteriophage phi-X174 (5386 bp ss-DNA) ss dna single stranded deoxyribonucleic acid ss DNA ss dna bpphx Enterobacteria phage phiX174 Bacteriophage phi-X174 This is the complete circular ssDNA genome of bacteriophage phi-X174 (5386 bp). dpra_bacsu DNA processing protein DprA dprA Protein Smf smf BSU16110 protein protein bacsu Bacillus subtilis MDQAAVCLTICRINQLLSPSLLLKWWKADPSMSLTSPVLQTVTRDQIKAAALKNEIEQFYPKLPRVLAAYREQGINTIPISSKQYPFWLKSIYDPPAVLFAKGDMTLLSKGRKIGIVGTRNPTAYGKQVVNHLTKEICRKGWVIVSGLASGIDGMSHAASIKAKGRTIGVIAGGFQHIYPRENLQLADHMAKHHILLSEHPPETKPQKWHFPMRNRIISGLSEGVIVVQGKEKSGSLITAYQALEQGREVFAVPGSLFDPYAGGPIKLIQQGAKAIWSAEDIFEELPERNVQYTEPF reca_bacsu Protein RecA recA recE BSU16940 Recombinase A protein protein bacsu Bacillus subtilis MSDRQAALDMALKQIEKQFGKGSIMKLGEKTDTRISTVPSGSLALDTALGIGGYPRGRIIEVYGPESSGKTTVALHAIAEVQQQGGQAAFIDAEHALDPVYAQKLGVNIEELLLSQPDTGEQALEIAEALVRSGAVDIVVVDSVAALVPKAEIEGDMGDSHVGLQARLMSQALRKLSGAINKSKTIAIFINQIREKVGVMFGNPETTPGGRALKFYSSVRLEVRRAEQLKQGNDVMGNKTKIKVVKNKVAPPFRTAEVDIMYGEGISKEGEIIDLGTELDIVQKSGSWYSYEEERLGQGRENAKQFLKENKDIMLMIQEQIREHYGLDNNGVVQQQAEETQEELEFEE reca_strpn Protein RecA recA SP_1940 Recombinase A protein protein strpn Streptococcus pneumoniae serotype 4 (strain ATCC BAA-334 / TIGR4) MAKKPKKLEEISKKFGAEREKALNDALKLIEKDFGKGSIMRLGERAEQKVQVMSSGSLALDIALGSGGYPKGRIIEIYGPESSGKTTVALHAVAQAQKEGGIAAFIDAEHALDPAYAAALGVNIDELLLSQPDSGEQGLEIAGKLIDSGAVDLVVVDSVAALVPRAEIDGDIGDSHVGLQARMMSQAMRKLGASINKTKTIAIFINQLREKVGVMFGNPETTPGGRALKFYASVRLDVRGNTQIKGTGDQKETNVGKETKIKVVKNKVAPPFKEAVVEIMYGEGISKTGELLKIASDLDIIKKAGAWYSYKDEKIGQGSENAKKYLAEHPEIFDEIDKQVRSKFGLIDGEEVSEQDTENKKDEPKKEEAVNEEVPLDLGDELEIEIEE 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) reca_ecoli Protein RecA recA lexB recH rnmB umuB tif zab b2699 Recombinase A JW2669 protein protein ecoli Escherichia coli (strain K12) MAIDENKQKALAAALGQIEKQFGKGSIMRLGEDRSMDVETISTGSLSLDIALGAGGLPMGRIVEIYGPESSGKTTLTLQVIAAAQREGKTCAFIDAEHALDPIYARKLGVDIDNLLCSQPDTGEQALEICDALARSGAVDVIVVDSVAALTPKAEIEGEIGDSHMGLAARMMSQAMRKLAGNLKQSNTLLIFINQIRMKIGVMFGNPETTTGGNALKFYASVRLDIRRIGAVKEGENVVGSETRVKVVKNKIAAPFKQAEFQILYGEGINFYGELVDLGVKEKLIEKAGAWYSYKGEKIGQGKANATAWLKDNPETAKEIEKKVRELLLSNPNSTPDFSVDDSEGVAETNEDF dt90_dna-q97qfo DprA interacts with a 90-mer poly-dT (dT90) 668404 N/A 668408 unspecified role unspecified role neutral component neutral component N/A 668409 unspecified role unspecified role neutral component neutral component direct interaction direct interaction false false false 2 B The 90-mer poly-dT was labelled with 32P. This indicates that DprA binds to nonspecific ssDNA. DprA was found to render dT90 resistant to a 3->5 ssDNA exonuclease. 80mer_rando-q97qfo-1 DprA interacts with an 80-mer of random sequence (ssDNA) 668404 N/A 668408 unspecified role unspecified role neutral component neutral component N/A 668410 unspecified role unspecified role neutral component neutral component direct interaction direct interaction false false false S3 A The 80-mer oligonucleotide of random sequence was labelled with 32P. This indicates that DprA binds to non-specific ssDNA. q97qf0-gen_bpphx DprA interacts with the complete genome of bacteriophage phi-X174 668404 N/A 668411 unspecified role unspecified role neutral component neutral component N/A 668408 unspecified role unspecified role neutral component neutral component direct interaction direct interaction false false false 3 A This indicates that a free end on the substrate oligonucleotide is not necessary for DprA binding. DprA also fully protected the phiX genome from Mung Bean nuclease. smf-gen_bpphx-1 Smf interacts with the complete genome of bacteriophage phi-X174 668404 N/A 668412 unspecified role unspecified role neutral component neutral component N/A 668411 unspecified role unspecified role neutral component neutral component direct interaction direct interaction false false false 3 A This indicates that a free end on the substrate oligonucleotide is not necessary for Smf binding. smf-smf Yeast two-hybrid assay reveals Smf self-interaction 668405 N/A 668412 unspecified role unspecified role bait bait N/A 668412 unspecified role unspecified role prey prey physical association physical association false false false S2 C q97qf0-q97qf0 Yeast two-hybrid assay reveals DprA self-interaction 668405 N/A 668408 unspecified role unspecified role bait bait N/A 668408 unspecified role unspecified role prey prey physical association physical association false false false S2 C smf-reca Yeast two-hybrid assay reveals Smf interacts with BsRecA 668405 N/A 668413 unspecified role unspecified role prey prey N/A 668412 unspecified role unspecified role bait bait physical association physical association false false false 5 B smf-reca-1 Yeast two-hybrid assay reveals DprA interacts with SpRecADeltaN 668405 N/A 668413 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 false N/A 668408 unspecified role unspecified role prey prey physical association physical association false false false 5 B reca-q97qf0 DprA copurifies with RecA from competent S. pneumonia cells 668406 N/A 668408 unspecified role unspecified role prey prey N/A 668414 unspecified role unspecified role bait bait c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-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 5 A reca-atp RecA has ssDNA-dependent ATPase activity 668407 N/A 668411 stimulator stimulator unspecified role unspecified role N/A 668415 enzyme target enzyme target substrate substrate unspecified role unspecified role N/A 668416 enzyme enzyme unspecified role unspecified role enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction false false false S4 A The ATP used was labelled with 32P ([32P]gammaATP). SSB - SSB has a strong inhibitory effect on the ssDNA-dependent ATPase activity of EcRecA. DprA - DprA has a stimulatory effect on the ssDNA-dependent ATPase activity of EcRecA.