MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. EMBO J. (0261-4189) 2011 imex curation Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. EMBO J. (0261-4189) 2011 imex curation Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. Only protein-protein interactions in vitro In vitro fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. EMBO J. (0261-4189) 2011 imex curation Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. Only protein-protein interactions in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. Structural biochemistry of nuclear actin-related proteins 4 and 8 reveals their interaction with actin. EMBO J. (0261-4189) 2011 imex curation Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. Only protein-protein interactions in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Fenn S., Breitsprecher D., Gerhold CB., Witte G., Faix J., Hopfner KP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation arp8_yeast Actin-like protein ARP8 ARP8 YOR3348C YOR141C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQEEAESSIIYEEPIDIPLEDDDDEDELEEENSVPLSSQADQENAENESDDSVDNVVGSETPRSVTGLSVDPRDVADEEDEDEEGEDEDEDEDDNDVDNEDENDNDNANENENELGSSRDKRAPPAVQTSKRYKKYPKLDPAKAPPGKKVPLHLLEKRRLGRIKAAEEFAKTLKKIGIEKVETTTLPATGLFQPLMLINQKNYSSDYLKKDDQIFALRDRKFLRNNNTSQISSTNTPDVIDLKSLPHSEASAAPLNDEIDLNDPTATIVIHPGSNSIKIGFPKDDHPVVVPNCVAVPKKWLDLENSEHVENVCLQREQSEEFNNIKSEMEKNFRERMRYYKRKVPGNAHEQVVSFNENSKPEIISEKNDPSPIEWIFDDSKLYYGSDALRCVDEKFVIRKPFRGGSFNVKSPYYKSLAELISDVTKLLEHALNSETLNVKPTKFNQYKVVLVIPDIFKKSHVETFIRVLLTELQFQAVAIIQESLATCYGAGISTSTCVVNIGAAETRIACVDEGTVLEHSAITLDYGGDDITRLFALFLLQSDFPLQDWKIDSKHGWLLAERLKKNFTTFQDADVAVQLYNFMNRSPNQPTEKYEFKLFDEVMLAPLALFFPQIFKLIRTSSHKNSSLEFQLPESRDLFTNELNDWNSLSQFESKEGNLYCDLNDDLKILNRILDAHNIIDQLQDKPENYGNTLKENFAPLEKAIVQSIANASITADVTRMNSFYSNILIVGGSSKIPALDFILTDRINIWRPSLLSSASFPQFYKKLTKEIKDLEGHYVNAPDKTEDENKQILQAQIKEKIVEELEEQHQNIEHQNGNEHIFPVSIIPPPRDMNPALIIWKGASVLAQIKLVEELFITNSDWDVHGSRILQYKCIFTY 8174851B6B077A19 8174851B6B077A19 acts_rabit Actin, alpha skeletal muscle ACTA1 ACTA Alpha-actin-1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MCDEDETTALVCDNGSGLVKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGDEAQSKRGILTLKYPIEHGIITNWDDMEKIWHHTFYNELRVAPEEHPTLLTEAPLNPKANREKMTQIMFETFNVPAMYVAIQAVLSLYASGRTTGIVLDSGDGVTHNVPIYEGYALPHAIMRLDLAGRDLTDYLMKILTERGYSFVTTAEREIVRDIKEKLCYVALDFENEMATAASSSSLEKSYELPDGQVITIGNERFRCPETLFQPSFIGMESAGIHETTYNSIMKCDIDIRKDLYANNVMSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWITKQEYDEAGPSIVHRKCF DIP protein P68135 original sequence version: 41 DIP protein P68135 original sequence version: 27 DIP protein P68135 original sequence version: 28 DIP protein P68135 original sequence version: 96 DIP protein P68135 original sequence version: 29 DIP protein P68135 original sequence version: 89 DIP protein P68135 original sequence version: 39 DIP protein P68135 original sequence version: 42 DIP protein P68135 original sequence version: 59 DIP protein P68135 original sequence version: 55 DIP protein P68135 original sequence version: 60 DIP protein P68135 original sequence version: 65 DIP protein P68135 original sequence version: 67 DIP protein P68135 original sequence version: 63 DIP protein P68135 original sequence version: 68 DIP protein P68135 original sequence version: 79 DIP protein P68135 original sequence version: 88 DIP protein P68135 original sequence version: 84 DIP protein P68135 original sequence version: 101 DIP protein P68135 original sequence version: 94 DIP protein P68135 original sequence version: 95 DIP protein P68135 original sequence version: 97 DIP protein P68135 original sequence version: 100 DIP protein P68135 original sequence version: 102 DIP protein P68135 original sequence version: 83 DF2A3A046346A179 arp4_yeast Actin-related protein 4 ARP4 ACT3 YJL081C J1012 Actin-like protein ARP4 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSNAALQVYGGDEVSAVVIDPGSYTTNIGYSGSDFPQSILPSVYGKYTADEGNKKIFSEQSIGIPRKDYELKPIIENGLVIDWDTAQEQWQWALQNELYLNSNSGIPALLTEPVWNSTENRKKSLEVLLEGMQFEACYLAPTSTCVSFAAGRPNCLVVDIGHDTCSVSPIVDGMTLSKSTRRNFIAGKFINHLIKKALEPKEIIPLFAIKQRKPEFIKKTFDYEVDKSLYDYANNRGFFQECKETLCHICPTKTLEETKTELSSTAKRSIESPWNEEIVFDNETRYGFAEELFLPKEDDIPANWPRSNSGVVKTWRNDYVPLKRTKPSGVNKSDKKVTPTEEKEQEAVSKSTSPAANSADTPNETGKRPLEEEKPPKENNELIGLADLVYSSIMSSDVDLRATLAHNVVLTGGTSSIPGLSDRLMTELNKILPSLKFRILTTGHTIERQYQSWLGGSILTSLGTFHQLWVGKKEYEEVGVERLLNDRFR 6DE1C8938C6C3DBB 6DE1C8938C6C3DBB acta1-acta1 4 Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction mint f5a f5b f5c f7a f7b f8a f8b we analysed the spontaneous polymerization of monomeric, pyrene-labelled Mg2þ-ATP-actin in the presence of Arp4 by measuring the increase in pyrene fluorescence. Arp4 decreased the polymerization rate of actin in a concentration- dependent manner (Figure 5A and B). However, it did not markedly sequester actin monomers at steady state, resulting in nearly constant levels of F-actin after 16 h (Figure 5C). Arp8 did not affect the polymerization kinetics of ATP-actin in pyrene assays (Figure 7A and B).Arp8 enhanced the inhibitory effect of Arp4 on actin polymerization in a concentration-dependent manner by as much as three-fold (Figure 8A and B). acta1-arp4 1 Arp4 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified direct interaction direct interaction mint sf5 acta1-arp8 1 Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified Arp8 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified direct interaction direct interaction mint sf7 SPR experiments to verify the direct interaction of Arp8 with actin indicate that Arp8 indeed binds to actin (Supplementary Figure S7). Due to problems with some unspecific binding to the reference channels of the chips, however, we were not able to determine an accurate KD. acta1-acta1-1 3 Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f5d mint Co-sedimentation analyses with F-actin and Arp4 confirmed that Arp4 does not reduce the amount of actin filaments, and moreover showed that Arp4 does not bind to actin filaments (Figure 5D). acta1-arp8-1 3 Arp8 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction mint Unexpectedly, and in contrast to Arp4, however, we found that Arp8 efficiently sequestered actin monomers at steady state, as assessed by fluorescence measurements and high-speed sedimentation analyses (Figure 7D and E). Co-sedimentation assays of 5 mM actin polymerized in the presence of the Arp4 and Arp8 concentrations indicated. The sequestration effect of Arp8 is more pronounced in the presence of Arp4 (compare with Figure 7E);(Figure 8D and E). f7d f7e f8d f8e acta1-acta1-2 2 Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Actin 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f5e mint we employed in vitro total internal reflection fluorescence (TIRF) microscopy on Oregon-Green (OG)-labelled, polymerizing actin filaments. We found that Arp4 decreased both the elongation rate of actin filaments as well as the total number of spontaneously formed filaments (Figure 5E).