A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex
All six minichromosome maintenance (MCM) proteins have DNA-dependent ATPase motifs in the central domain which is conserved from yeast to mammals. Our group purified MCM protein complexes consisting of MCM2, -4 (Cdc21), -6 (Mis5), and -7 (CDC47) proteins from HeLa cells by using histone-Sepharose co...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 1997-09, Vol.272 (39), p.24508-24513 |
|---|---|
| 1. Verfasser: | |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 24513 |
|---|---|
| container_issue | 39 |
| container_start_page | 24508 |
| container_title | The Journal of biological chemistry |
| container_volume | 272 |
| creator | Ishimi, Yukio |
| description | All six minichromosome maintenance (MCM) proteins have DNA-dependent ATPase motifs in the central domain which is conserved from yeast to mammals. Our group purified MCM protein complexes consisting of MCM2, -4 (Cdc21), -6 (Mis5), and -7 (CDC47) proteins from HeLa cells by using histone-Sepharose column chromatography (Ishimi, Y., Ichinose, S., Omori, A., Sato K., and Kimura, H. (1996) J. Biol. Chem. 271, 24115–24122). The present study revealed that both ATPase activity and DNA helicase activity that displaces oligonucleotides annealed to single-stranded circular DNA are associated with an MCM protein complex. Both ATPase and DNA helicase activities were co-purified with a 600-kDa protein complex that is consisted of equal amounts of MCM4, -6, and -7 proteins. An immunodepletion of the MCM protein complex from the purified fraction using anti-MCM4 antibody resulted in the severe reduction of the DNA helicase activity. Displacement of DNA fragments by the DNA helicase suggested that it migrated along single-stranded DNA in the 3′ to 5′ direction, and the DNA helicase activity was detected only in the presence of hydrolyzable ATP or dATP. These results suggest that this helicase may be involved in the initiation of DNA replication as a DNA unwinding enzyme. |
| doi_str_mv | 10.1074/jbc.272.39.24508 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_16098914</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925819636276</els_id><sourcerecordid>16098914</sourcerecordid><originalsourceid>FETCH-LOGICAL-c416t-2259b323608f0de29eb60498f0d7b0a6dbfa70981c1e5b6451e11faf84ce8cef3</originalsourceid><addsrcrecordid>eNp1kM9LwzAUx4Moc07vXoQcxNNak_RnvJX6YwOnHhS8hTR9tRntOptuc_-9mR0eBAOP8Hjf75f3PgidU-JSEvnX80y5LGKux13mByQ-QENKYs_xAvp-iIaEMOpwFsTH6MSYObHP53SABtwjAaf-ED0l-NbWBCqtpAGcqE6vdbfFU4MTYxqlZQc53uiuxHKBZ-nMH2MnHNsmx06EX9qmA73AaVMvK_g6RUeFrAyc7f8Reru_e00nzuPzwzRNHh3l07BzGAt45jEvJHFBcmAcstButmuijMgwzwoZER5TRSHIQj-gQGkhi9hXECsovBG66nOXbfO5AtOJWhsFVSUX0KyMoKF12wOtkPRC1TbGtFCIZatr2W4FJWKHUFiEwiIUHhc_CK3lYp-9ymrIfw17ZnZ-2c9L_VFudAsi040qof4bc9PLwHJYa2iFURoWCnJrUZ3IG_3_Dt9Yh4jD</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16098914</pqid></control><display><type>article</type><title>A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Ishimi, Yukio</creator><creatorcontrib>Ishimi, Yukio</creatorcontrib><description>All six minichromosome maintenance (MCM) proteins have DNA-dependent ATPase motifs in the central domain which is conserved from yeast to mammals. Our group purified MCM protein complexes consisting of MCM2, -4 (Cdc21), -6 (Mis5), and -7 (CDC47) proteins from HeLa cells by using histone-Sepharose column chromatography (Ishimi, Y., Ichinose, S., Omori, A., Sato K., and Kimura, H. (1996) J. Biol. Chem. 271, 24115–24122). The present study revealed that both ATPase activity and DNA helicase activity that displaces oligonucleotides annealed to single-stranded circular DNA are associated with an MCM protein complex. Both ATPase and DNA helicase activities were co-purified with a 600-kDa protein complex that is consisted of equal amounts of MCM4, -6, and -7 proteins. An immunodepletion of the MCM protein complex from the purified fraction using anti-MCM4 antibody resulted in the severe reduction of the DNA helicase activity. Displacement of DNA fragments by the DNA helicase suggested that it migrated along single-stranded DNA in the 3′ to 5′ direction, and the DNA helicase activity was detected only in the presence of hydrolyzable ATP or dATP. These results suggest that this helicase may be involved in the initiation of DNA replication as a DNA unwinding enzyme.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.272.39.24508</identifier><identifier>PMID: 9305914</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Adenosine Triphosphatases - metabolism ; Biological Transport ; Cell Cycle Proteins - metabolism ; DNA Helicases - metabolism ; DNA-Binding Proteins - metabolism ; Fungal Proteins - metabolism ; HeLa Cells ; Humans ; Minichromosome Maintenance Complex Component 4 ; Minichromosome Maintenance Complex Component 6 ; Minichromosome Maintenance Complex Component 7 ; Nuclear Proteins - metabolism ; Protein Binding ; Schizosaccharomyces pombe Proteins</subject><ispartof>The Journal of biological chemistry, 1997-09, Vol.272 (39), p.24508-24513</ispartof><rights>1997 © 1997 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c416t-2259b323608f0de29eb60498f0d7b0a6dbfa70981c1e5b6451e11faf84ce8cef3</citedby><cites>FETCH-LOGICAL-c416t-2259b323608f0de29eb60498f0d7b0a6dbfa70981c1e5b6451e11faf84ce8cef3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9305914$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ishimi, Yukio</creatorcontrib><title>A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>All six minichromosome maintenance (MCM) proteins have DNA-dependent ATPase motifs in the central domain which is conserved from yeast to mammals. Our group purified MCM protein complexes consisting of MCM2, -4 (Cdc21), -6 (Mis5), and -7 (CDC47) proteins from HeLa cells by using histone-Sepharose column chromatography (Ishimi, Y., Ichinose, S., Omori, A., Sato K., and Kimura, H. (1996) J. Biol. Chem. 271, 24115–24122). The present study revealed that both ATPase activity and DNA helicase activity that displaces oligonucleotides annealed to single-stranded circular DNA are associated with an MCM protein complex. Both ATPase and DNA helicase activities were co-purified with a 600-kDa protein complex that is consisted of equal amounts of MCM4, -6, and -7 proteins. An immunodepletion of the MCM protein complex from the purified fraction using anti-MCM4 antibody resulted in the severe reduction of the DNA helicase activity. Displacement of DNA fragments by the DNA helicase suggested that it migrated along single-stranded DNA in the 3′ to 5′ direction, and the DNA helicase activity was detected only in the presence of hydrolyzable ATP or dATP. These results suggest that this helicase may be involved in the initiation of DNA replication as a DNA unwinding enzyme.</description><subject>Adenosine Triphosphatases - metabolism</subject><subject>Biological Transport</subject><subject>Cell Cycle Proteins - metabolism</subject><subject>DNA Helicases - metabolism</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Fungal Proteins - metabolism</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>Minichromosome Maintenance Complex Component 4</subject><subject>Minichromosome Maintenance Complex Component 6</subject><subject>Minichromosome Maintenance Complex Component 7</subject><subject>Nuclear Proteins - metabolism</subject><subject>Protein Binding</subject><subject>Schizosaccharomyces pombe Proteins</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM9LwzAUx4Moc07vXoQcxNNak_RnvJX6YwOnHhS8hTR9tRntOptuc_-9mR0eBAOP8Hjf75f3PgidU-JSEvnX80y5LGKux13mByQ-QENKYs_xAvp-iIaEMOpwFsTH6MSYObHP53SABtwjAaf-ED0l-NbWBCqtpAGcqE6vdbfFU4MTYxqlZQc53uiuxHKBZ-nMH2MnHNsmx06EX9qmA73AaVMvK_g6RUeFrAyc7f8Reru_e00nzuPzwzRNHh3l07BzGAt45jEvJHFBcmAcstButmuijMgwzwoZER5TRSHIQj-gQGkhi9hXECsovBG66nOXbfO5AtOJWhsFVSUX0KyMoKF12wOtkPRC1TbGtFCIZatr2W4FJWKHUFiEwiIUHhc_CK3lYp-9ymrIfw17ZnZ-2c9L_VFudAsi040qof4bc9PLwHJYa2iFURoWCnJrUZ3IG_3_Dt9Yh4jD</recordid><startdate>19970926</startdate><enddate>19970926</enddate><creator>Ishimi, Yukio</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope></search><sort><creationdate>19970926</creationdate><title>A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex</title><author>Ishimi, Yukio</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c416t-2259b323608f0de29eb60498f0d7b0a6dbfa70981c1e5b6451e11faf84ce8cef3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Adenosine Triphosphatases - metabolism</topic><topic>Biological Transport</topic><topic>Cell Cycle Proteins - metabolism</topic><topic>DNA Helicases - metabolism</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Fungal Proteins - metabolism</topic><topic>HeLa Cells</topic><topic>Humans</topic><topic>Minichromosome Maintenance Complex Component 4</topic><topic>Minichromosome Maintenance Complex Component 6</topic><topic>Minichromosome Maintenance Complex Component 7</topic><topic>Nuclear Proteins - metabolism</topic><topic>Protein Binding</topic><topic>Schizosaccharomyces pombe Proteins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ishimi, Yukio</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ishimi, Yukio</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1997-09-26</date><risdate>1997</risdate><volume>272</volume><issue>39</issue><spage>24508</spage><epage>24513</epage><pages>24508-24513</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>All six minichromosome maintenance (MCM) proteins have DNA-dependent ATPase motifs in the central domain which is conserved from yeast to mammals. Our group purified MCM protein complexes consisting of MCM2, -4 (Cdc21), -6 (Mis5), and -7 (CDC47) proteins from HeLa cells by using histone-Sepharose column chromatography (Ishimi, Y., Ichinose, S., Omori, A., Sato K., and Kimura, H. (1996) J. Biol. Chem. 271, 24115–24122). The present study revealed that both ATPase activity and DNA helicase activity that displaces oligonucleotides annealed to single-stranded circular DNA are associated with an MCM protein complex. Both ATPase and DNA helicase activities were co-purified with a 600-kDa protein complex that is consisted of equal amounts of MCM4, -6, and -7 proteins. An immunodepletion of the MCM protein complex from the purified fraction using anti-MCM4 antibody resulted in the severe reduction of the DNA helicase activity. Displacement of DNA fragments by the DNA helicase suggested that it migrated along single-stranded DNA in the 3′ to 5′ direction, and the DNA helicase activity was detected only in the presence of hydrolyzable ATP or dATP. These results suggest that this helicase may be involved in the initiation of DNA replication as a DNA unwinding enzyme.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>9305914</pmid><doi>10.1074/jbc.272.39.24508</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 1997-09, Vol.272 (39), p.24508-24513 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16098914 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Adenosine Triphosphatases - metabolism Biological Transport Cell Cycle Proteins - metabolism DNA Helicases - metabolism DNA-Binding Proteins - metabolism Fungal Proteins - metabolism HeLa Cells Humans Minichromosome Maintenance Complex Component 4 Minichromosome Maintenance Complex Component 6 Minichromosome Maintenance Complex Component 7 Nuclear Proteins - metabolism Protein Binding Schizosaccharomyces pombe Proteins |
| title | A DNA Helicase Activity Is Associated with an MCM4, -6, and -7 Protein Complex |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-13T04%3A58%3A43IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20DNA%20Helicase%20Activity%20Is%20Associated%20with%20an%20MCM4,%20-6,%20and%20-7%20Protein%20Complex&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Ishimi,%20Yukio&rft.date=1997-09-26&rft.volume=272&rft.issue=39&rft.spage=24508&rft.epage=24513&rft.pages=24508-24513&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.272.39.24508&rft_dat=%3Cproquest_cross%3E16098914%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16098914&rft_id=info:pmid/9305914&rft_els_id=S0021925819636276&rfr_iscdi=true |