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 GINS, a central nexus in the archaeal DNA replication fork. GINS, a central nexus in the archaeal DNA replication fork. EMBO Rep. (1469-221X) 2006 imex curation Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. 2009-01-08 Only protein-protein interactions in vitro In vitro pull down pull down nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation GINS, a central nexus in the archaeal DNA replication fork. GINS, a central nexus in the archaeal DNA replication fork. EMBO Rep. (1469-221X) 2006 imex curation Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. 2009-01-08 Only protein-protein interactions sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) chromatography chromatography technology column chromatography chromatography nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation GINS, a central nexus in the archaeal DNA replication fork. GINS, a central nexus in the archaeal DNA replication fork. EMBO Rep. (1469-221X) 2006 imex curation Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. 2009-01-08 Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation GINS, a central nexus in the archaeal DNA replication fork. GINS, a central nexus in the archaeal DNA replication fork. EMBO Rep. (1469-221X) 2006 imex curation Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. 2009-01-08 Only protein-protein interactions sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation GINS, a central nexus in the archaeal DNA replication fork. GINS, a central nexus in the archaeal DNA replication fork. EMBO Rep. (1469-221X) 2006 imex curation Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Marinsek N., Barry ER., Makarova KS., Dionne I., Koonin EV., Bell SD. EMBO Rep. (1469-221X) 2006 2009-01-08 Only protein-protein interactions imex curation mcm_sacs2 Minichromosome maintenance protein MCM MCM SSO0774 protein protein sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) MEIPSKQIDYRDVFIEFLTTFKGNNNQNKYIERINELVAYRKKSLIIEFSDVLSFNENLAYEIINNTKIILPILEGALYDHILQLDPTYQRDIEKVHVRIVGIPRVIELRKIRSTDIGKLITIDGILVKVTPVKERIYKATYKHIHPDCMQEFEWPEDEEMPEVLEMPTICPKCGKPGQFRLIPEKTKLIDWQKAVIQERPEEVPSGQLPRQLEIILEDDLVDSARPGDRVKVTGILDIKQDSPVKRGSRAVFDIYMKVSSIEVSQKVLDEVIISEEDEKKIKDLAKDPWIRDRIISSIAPSIYGHWELKEALALALFGGVPKVLEDTRIRGDIHILIIGDPGTAKSQMLQFISRVAPRAVYTTGKGSTAAGLTAAVVREKGTGEYYLEAGALVLADGGIAVIDEIDKMRDEDRVAIHEAMEQQTVSIAKAGIVAKLNARAAVIAAGNPKFGRYISERPVSDNINLPPTILSRFDLIFILKDQPGEQDRELANYILDVHSGKSTKNIIDIDTLRKYIAYARKYVTPKITSEAKNLITDFFVEMRKKSSETPDSPILITPRQLEALIRISEAYAKMALKAEVTREDAERAINIMRLFLESVGVDMESGKIDIDTIMTGKPKSAREKMMKIIEIIDSLAVSSECAKVKDILKEAQQVGIEKSNIEKLLTDMRKSGIIYEAKPECYKKV DIP protein Q9UXG1 original sequence version: 74 mint Has ATPase and DNA helicase activities. The latter preferentially melts 5'-tailed oligonucleotides and is stimulated by the SSB protein (single-stranded DNA binding protein) C3ECD380F272E77E q980j6_sacs2 SSO0295 protein protein sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) MVKRNSKEIVMEVFLGEPNNQEIREVVRNVYSKDNLCLSAYYSIEPILFSFIISKNTKNSFTLSFIPEECETQIRLDANGKWIIDNKKKYFLGQNSFCSYLPITAEDVLPIVSCIITSTILDRRSLSEWELETIREMEKFDMTIEKNLKIPAYKTLPAFLSLMFSLDPYIPGISGNRENAISLIREVNASEISKLEDLNESQLNTLLFRIISTIVKENPKFTRDDIITDRFFYLDYDILELAFSLIYSFDTIGSNEILQFVLSSSYAEALISRFRQTLSKGFSINSIENKQSYYIVETSNLTSPLLAQIILLQLQKIRRDKMIVIRDNDRLFTSRFFIPSVKKEGLIKIDDRVKNLIGI mint CEB3950D2BF44F7F q97z82_sacs2 SSO1049 protein protein sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) MLDELVKKELSEEEITEIKLEEIIKYITLIKKSKTFVSSEIRKEELKFLSELAESLFELRLSKVLEGKVGKGFDEFIFDIFKILKQFYVDLLTGRYIIYNDKIYCIVQKPLIYNDHRVNEGDVLVLPMREALPLIIASYLTPYKIDIEEQL mint 857E458321AEE4F8 pris_sacs2 DNA primase small subunit PriS priS priA SSO1048 protein protein sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) MGTFTLHQGQTNLIKSFFRNYYLNAELELPKDMELREFALQPFGSDTYVRHLSFSSSEELRDYLVNRNLPLHLFYSSARYQLPSARNMEEKAWMGSDLLFDIDADHLCKLRSIRFCPVCGNAVVSEKCERDNVETLEYVEMTSECIKRGLEQTRNLVEILEDDFGLKPKVYFSGNRGFHVQVDCYGNCALLDSDERKEIAEYVMGIGVPGYPGGSENAPGWVGRKNRGINGVTIDEQVTIDVKRLIRIPNSLHGKSGLIVKRVPNLDDFEFNETLSPFTGYTIFLPYITIETEVLGSIIKLNRGIPIKIKSSIGIYLHLRNLGEVKAYVR DIP protein Q97Z83 original sequence version: 66 mint DNA primase is the polymerase that synthesizes small RNA primers for the Okazaki fragments on both template strands at replication forks during chromosomal DNA synthesis E9B3C556E619B66E q9uxg0_sacso ORF-c40_009 SSOP1_0803 protein protein sulsf Sulfolobus solfataricus MGSATSNIRFCIKKKYIMIEVKLRAIKRLSNVYTRRVMIIEDWNGSSITTGNIELVKGSENQLPQWLAIILEGKKVAKIEDKISIEDLGRILFQERQNMNTPASLVPLGKDFTSRVQLYLETLRKDNNVESLEKLRKSIGILNEIIKIRLRKLIQLAFLNIDDQNLINGMTEEELLIYKTIKQLIKELYGDIIGNS mint 468FEA66643FE88F pril_sacs2 DNA primase large subunit PriL priL priB C21_042 SSO0557 protein protein sulso Sulfolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2) MALDVKKYPFIKSLDDELKKYGGGITLTDLLLNSTTLIDQAKDRIQKTKSGDELPHYVSYNEPVLVFYTTLLSLAILNDVKLIRRYAYAEAKQFRSLLHTENEENLLEISKLLDLKINRCDPIKFYLEKKRRIIQKEFCVHFIDYLKYTKDLKEDWKLSGQILHKGYVYLDKNQLIGLIAESIKSKIVEMIRPLNLKEIPEKLKSLIERRGIIPPCIENILAKEKLNEEEIRTLITFYIDIGKGLSGIVSIMKKYNVSNVEDLYRKYRGDKGTRYIVYSCAKMKQLGLCVSSCNVKNPLQLYFLSNE DIP protein Q9UWW1 original sequence version: 71 mint DNA primase is the polymerase that synthesizes small RNA primers for the Okazaki fragments on both template strands at replication forks during chromosomal DNA synthesis 5EF09A913B4ABAC3 orf_c40_009-mcm 5 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F1A EMBO-Reports In the course of a two-hybrid screen to identify potential binding partners of the MCM protein of the hyperthermophilic archaeon Sulfolobus solfataricus, we identified open reading frame SSO0772 as a candidate interaction partner.Yeast cells were transformed with these constructs together with MCM-DBD- and MCM-AD-expressing plasmids. As can be seen in Fig 1A, the SSO0772 protein interacts with MCM in both DBD and AD fusion contexts. Next, we tested the ability of SSO0772 to interact with amino- and carboxy-terminally truncated MCM. The results shown in Fig 1B indicate that SSO0772 interacts with the N-terminal 266 residues of MCM. mint mcm-orf_c40_009 5 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain range MINT-1896129: original sequence: 1-266 : MEIPSKQIDYRDVFIEFLTTFKGNNNQNKYIERINELVAYRKKSLIIEFSDVLSFNENLAYEIINNTKIILPILEGALYDHILQLDPTYQRDIEKVHVRIVGIPRVIELRKIRSTDIGKLITIDGILVKVTPVKERIYKATYKHIHPDCMQEFEWPEDEEMPEVLEMPTICPKCGKPGQFRLIPEKTKLIDWQKAVIQERPEEVPSGQLPRQLEIILEDDLVDSARPGDRVKVTGILDIKQDSPVKRGSRAVFDIYMKVSSIEVS Stoichiometry: 1.0 physical association physical association F1A EMBO-Reports In the course of a two-hybrid screen to identify potential binding partners of the MCM protein of the hyperthermophilic archaeon Sulfolobus solfataricus, we identified open reading frame SSO0772 as a candidate interaction partner.Yeast cells were transformed with these constructs together with MCM-DBD- and MCM-AD-expressing plasmids. As can be seen in Fig 1A, the SSO0772 protein interacts with MCM in both DBD and AD fusion contexts. Next, we tested the ability of SSO0772 to interact with amino- and carboxy-terminally truncated MCM. The results shown in Fig 1B indicate that SSO0772 interacts with the N-terminal 266 residues of MCM. mint orf_c40_009-q97z82 5 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports A two-hybrid matrix indicated that Gins15 could interact with itself, with Gins23 and with RecJdbh. mint orf_c40_009-mcm-1 5 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports As described above, Gins23 interacted with MCM and also with both large and small subunits of the archaeal primase (Fig 4A) mint orf_c40_009-prib 5 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports As described above, Gins23 interacted with MCM and also with both large and small subunits of the archaeal primase (Fig 4A) mint orf_c40_009-pria 5 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports As described above, Gins23 interacted with MCM and also with both large and small subunits of the archaeal primase (Fig 4A) mint q97z82-orf_c40_009 5 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports A two-hybrid matrix indicated that Gins15 could interact with itself, with Gins23 and with RecJdbh. mint q97z82-q97z82 5 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports A two-hybrid matrix indicated that Gins15 could interact with itself, with Gins23 and with RecJdbh. mint q97z82-q980j6 5 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports A two-hybrid matrix indicated that Gins15 could interact with itself, with Gins23 and with RecJdbh. mint q980j6-q97z82 5 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports A two-hybrid matrix indicated that Gins15 could interact with itself, with Gins23 and with RecJdbh. mint mcm-mcm 5 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports mint mcm-orf_c40_009-1 5 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports As described above, Gins23 interacted with MCM and also with both large and small subunits of the archaeal primase (Fig 4A) mint prib-orf_c40_009 5 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports As described above, Gins23 interacted with MCM and also with both large and small subunits of the archaeal primase (Fig 4A) mint prib-pria 5 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports mint pria-prib-1 5 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports mint pria-pria 5 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports mint pria-orf_c40_009 5 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell tagged molecule tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F4A EMBO-Reports As described above, Gins23 interacted with MCM and also with both large and small subunits of the archaeal primase (Fig 4A) mint orf_c40_009-mcm-2 4 6 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 10 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2D F2C EMBO-Reports To test whether the MCM Gins23 interaction detected in the yeast two-hybrid assay was representative of a physiologically relevant interaction in S. solfataricus cells, we carried out immunoprecipitation assays. The results indicate that Gins23 and MCM can be co-immunoprecipitated. We tested the ability of MCM and Gins23 to co-precipitate from extracts prepared from cells in early and mid-logarithmic growth and also from stationary phase. mint orf_c40_009-q97z82-1 4 8 fingerprinting peptide massfingerprinting fingerprinting predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 10 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F3A EMBO-Reports As can be seen in Fig 3A, three proteins eluted specifically from the anti-Gins23 antisera column. These were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry as Gins23 and the products of the SSO0295 and SSO1049 open reading frames. mint orf_c40_009-q980j6 4 10 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 fingerprinting peptide massfingerprinting fingerprinting predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F3A EMBO-Reports As can be seen in Fig 3A, three proteins eluted specifically from the anti-Gins23 antisera column. These were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry as Gins23 and the products of the SSO0295 and SSO1049 open reading frames. mint orf_c40_00-orf_c40_0 1 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified his tagged his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 direct interaction direct interaction F4C EMBO-Reports As can be seen in the upper panels of Fig 4C,D, GINS was retained on columns containing MCM or primase, but not by the matrix alone, or by matrix containing PCNA2 mint prib-orf_c40_009-1 1 11 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified his tagged his tagged his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 direct interaction direct interaction F4D EMBO-Reports As can be seen in the upper panels of Fig 4C,D, GINS was retained on columns containing MCM or primase, but not by the matrix alone, or by matrix containing PCNA2 mint orf_c40_009-q980j6-1 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 colocalization colocalization F3E EMBO-Reports As can been seen in Fig 3E, the two GINS family members and RecJdbh co-purified in eight steps, supporting the notion that they exist as a stable complex in cells. mint q97z82-q980j6-1 2 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 colocalization colocalization F3E EMBO-Reports As can been seen in Fig 3E, the two GINS family members and RecJdbh co-purified in eight steps, supporting the notion that they exist as a stable complex in cells. mint orf_c40_009-q97z82-2 2 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 colocalization colocalization F3E EMBO-Reports As can been seen in Fig 3E, the two GINS family members and RecJdbh co-purified in eight steps, supporting the notion that they exist as a stable complex in cells. mint orf_c40_009-q97z82-3 3 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 1.0 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 1.0 direct interaction direct interaction F4B EMBO-Reports Analytical gel filtration gave an apparent molecular mass of 67 kDa. As a dimer of Gins23 and Gins15 would have a mass of 37 kDa, it seems that the core archaeal GINS complex is tetrameric, containing a dimer of dimers. mint