The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication
Human single-stranded DNA binding protein (human SSB) is a multisubunit protein containing polypeptides of 70, 34, and 11 kDa that is required for SV40 DNA replication in vitro. In this report we identify the functions of the SSB and its individual subunits in SV40 DNA replication. The 70 kDa subuni...
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| Veröffentlicht in: | The Journal of biological chemistry 1990-05, Vol.265 (13), p.7693-7700 |
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| creator | KENNY, M. K SCHLEGEL, U FURNEAUX, H HURWITZ, J |
| description | Human single-stranded DNA binding protein (human SSB) is a multisubunit protein containing polypeptides of 70, 34, and 11
kDa that is required for SV40 DNA replication in vitro. In this report we identify the functions of the SSB and its individual
subunits in SV40 DNA replication. The 70 kDa subunit was found to bind to single-stranded DNA, whereas the other subunits
did not. Four monoclonal antibodies against human SSB were isolated which inhibited SV40 DNA replication in vitro. The antibodies
have been designated alpha SSB70A, alpha SSB70B, alpha SSB70C, and alpha SSB34A to indicate which subunits are recognized.
Immunolocalization experiments indicated that human SSB is a nuclear protein. Human SSB is required for the SV40 large tumor
antigen-catalyzed unwinding of SV40 DNA and stimulates DNA polymerases (pol) alpha and delta. The DNA unwinding reaction and
stimulation of pol delta were blocked by alpha SSB70C, whereas the stimulation of pol alpha by human SSB was unaffected by
this antibody. Conversely, alpha SSB70A, -70B, and -34A inhibited the stimulation of pol alpha, but they had no effect on
DNA unwinding and pol delta stimulation. None of the antibodies inhibited the binding of SSB to single-stranded DNA. These
results suggest that DNA unwinding and stimulation of pol alpha and pol delta are required functions of human SSB in SV40
DNA replication. The human SSB 70-kDa subunit appears to be required for DNA unwinding and pol delta stimulation, whereas
both the 70- and 34-kDa subunits may be involved in the stimulation of pol alpha. |
| doi_str_mv | 10.1016/S0021-9258(19)39170-7 |
| format | Article |
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kDa that is required for SV40 DNA replication in vitro. In this report we identify the functions of the SSB and its individual
subunits in SV40 DNA replication. The 70 kDa subunit was found to bind to single-stranded DNA, whereas the other subunits
did not. Four monoclonal antibodies against human SSB were isolated which inhibited SV40 DNA replication in vitro. The antibodies
have been designated alpha SSB70A, alpha SSB70B, alpha SSB70C, and alpha SSB34A to indicate which subunits are recognized.
Immunolocalization experiments indicated that human SSB is a nuclear protein. Human SSB is required for the SV40 large tumor
antigen-catalyzed unwinding of SV40 DNA and stimulates DNA polymerases (pol) alpha and delta. The DNA unwinding reaction and
stimulation of pol delta were blocked by alpha SSB70C, whereas the stimulation of pol alpha by human SSB was unaffected by
this antibody. Conversely, alpha SSB70A, -70B, and -34A inhibited the stimulation of pol alpha, but they had no effect on
DNA unwinding and pol delta stimulation. None of the antibodies inhibited the binding of SSB to single-stranded DNA. These
results suggest that DNA unwinding and stimulation of pol alpha and pol delta are required functions of human SSB in SV40
DNA replication. The human SSB 70-kDa subunit appears to be required for DNA unwinding and pol delta stimulation, whereas
both the 70- and 34-kDa subunits may be involved in the stimulation of pol alpha.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(19)39170-7</identifier><identifier>PMID: 2159011</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Biochemistry and Molecular Biology</publisher><subject>Antibodies, Monoclonal ; Base Sequence ; Biological and medical sciences ; Cell Line ; DNA Probes ; DNA Replication ; DNA, Viral - genetics ; DNA-Binding Proteins - isolation & purification ; DNA-Binding Proteins - metabolism ; Fundamental and applied biological sciences. Psychology ; HeLa Cells - metabolism ; Humans ; Kinetics ; Macromolecular Substances ; Molecular and cellular biology ; Molecular genetics ; Molecular Sequence Data ; Molecular Weight ; Oligonucleotide Probes ; Replication ; Simian virus 40 - genetics</subject><ispartof>The Journal of biological chemistry, 1990-05, Vol.265 (13), p.7693-7700</ispartof><rights>1991 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c440t-4ba44877bef79a5302c431837361c55cd5c344874634e425b07eae06a7b634303</citedby><cites>FETCH-LOGICAL-c440t-4ba44877bef79a5302c431837361c55cd5c344874634e425b07eae06a7b634303</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=19838608$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2159011$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>KENNY, M. K</creatorcontrib><creatorcontrib>SCHLEGEL, U</creatorcontrib><creatorcontrib>FURNEAUX, H</creatorcontrib><creatorcontrib>HURWITZ, J</creatorcontrib><title>The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Human single-stranded DNA binding protein (human SSB) is a multisubunit protein containing polypeptides of 70, 34, and 11
kDa that is required for SV40 DNA replication in vitro. In this report we identify the functions of the SSB and its individual
subunits in SV40 DNA replication. The 70 kDa subunit was found to bind to single-stranded DNA, whereas the other subunits
did not. Four monoclonal antibodies against human SSB were isolated which inhibited SV40 DNA replication in vitro. The antibodies
have been designated alpha SSB70A, alpha SSB70B, alpha SSB70C, and alpha SSB34A to indicate which subunits are recognized.
Immunolocalization experiments indicated that human SSB is a nuclear protein. Human SSB is required for the SV40 large tumor
antigen-catalyzed unwinding of SV40 DNA and stimulates DNA polymerases (pol) alpha and delta. The DNA unwinding reaction and
stimulation of pol delta were blocked by alpha SSB70C, whereas the stimulation of pol alpha by human SSB was unaffected by
this antibody. Conversely, alpha SSB70A, -70B, and -34A inhibited the stimulation of pol alpha, but they had no effect on
DNA unwinding and pol delta stimulation. None of the antibodies inhibited the binding of SSB to single-stranded DNA. These
results suggest that DNA unwinding and stimulation of pol alpha and pol delta are required functions of human SSB in SV40
DNA replication. The human SSB 70-kDa subunit appears to be required for DNA unwinding and pol delta stimulation, whereas
both the 70- and 34-kDa subunits may be involved in the stimulation of pol alpha.</description><subject>Antibodies, Monoclonal</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>DNA Probes</subject><subject>DNA Replication</subject><subject>DNA, Viral - genetics</subject><subject>DNA-Binding Proteins - isolation & purification</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>HeLa Cells - metabolism</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Macromolecular Substances</subject><subject>Molecular and cellular biology</subject><subject>Molecular genetics</subject><subject>Molecular Sequence Data</subject><subject>Molecular Weight</subject><subject>Oligonucleotide Probes</subject><subject>Replication</subject><subject>Simian virus 40 - genetics</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1990</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtv1DAUhS0EKtOBn1DJEgLBIuAbv-JlVSitVMGCIrGzHOemMcpjsJOi_nucmVG7xBtL55z72bqHkDNgH4GB-vSDsRIKU8rqPZgP3IBmhX5GNsAqXnAJv56TzWPkJTlN6TfLRxg4ISclSMMANmS-7ZDGqUc6tbRbBjfSFMa7Hos0Rzc22NDP385pHcYmy3QXpxnDSLNDw5zoKt-HZnE9TUu9jActI4aQSfchLokKtkdE3PXBuzlM4yvyonV9wtfHe0t-Xn65vbgqbr5_vb44vym8EGwuRO2EqLSusdXGSc5KLzhUXHMFXkrfSM_XgFBcoChlzTQ6ZMrpOiuc8S15d-Dmb_9ZMM12CMlj37sRpyVZbbQwrDL_DYJUIJlaifIQ9HFKKWJrdzEMLj5YYHatxe5rsevOLRi7r8XqPHd2fGCpB2wep449ZP_t0XfJu77Nq_chPcFNxSuVi92SN4dcF-66vyGircPkOxxsqaQFbrUynP8D_vafpg</recordid><startdate>19900505</startdate><enddate>19900505</enddate><creator>KENNY, M. K</creator><creator>SCHLEGEL, U</creator><creator>FURNEAUX, H</creator><creator>HURWITZ, J</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>IQODW</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><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>19900505</creationdate><title>The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication</title><author>KENNY, M. K ; SCHLEGEL, U ; FURNEAUX, H ; HURWITZ, J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-4ba44877bef79a5302c431837361c55cd5c344874634e425b07eae06a7b634303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1990</creationdate><topic>Antibodies, Monoclonal</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>DNA Probes</topic><topic>DNA Replication</topic><topic>DNA, Viral - genetics</topic><topic>DNA-Binding Proteins - isolation & purification</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>HeLa Cells - metabolism</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Macromolecular Substances</topic><topic>Molecular and cellular biology</topic><topic>Molecular genetics</topic><topic>Molecular Sequence Data</topic><topic>Molecular Weight</topic><topic>Oligonucleotide Probes</topic><topic>Replication</topic><topic>Simian virus 40 - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>KENNY, M. K</creatorcontrib><creatorcontrib>SCHLEGEL, U</creatorcontrib><creatorcontrib>FURNEAUX, H</creatorcontrib><creatorcontrib>HURWITZ, J</creatorcontrib><collection>Pascal-Francis</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><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>KENNY, M. K</au><au>SCHLEGEL, U</au><au>FURNEAUX, H</au><au>HURWITZ, J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1990-05-05</date><risdate>1990</risdate><volume>265</volume><issue>13</issue><spage>7693</spage><epage>7700</epage><pages>7693-7700</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>Human single-stranded DNA binding protein (human SSB) is a multisubunit protein containing polypeptides of 70, 34, and 11
kDa that is required for SV40 DNA replication in vitro. In this report we identify the functions of the SSB and its individual
subunits in SV40 DNA replication. The 70 kDa subunit was found to bind to single-stranded DNA, whereas the other subunits
did not. Four monoclonal antibodies against human SSB were isolated which inhibited SV40 DNA replication in vitro. The antibodies
have been designated alpha SSB70A, alpha SSB70B, alpha SSB70C, and alpha SSB34A to indicate which subunits are recognized.
Immunolocalization experiments indicated that human SSB is a nuclear protein. Human SSB is required for the SV40 large tumor
antigen-catalyzed unwinding of SV40 DNA and stimulates DNA polymerases (pol) alpha and delta. The DNA unwinding reaction and
stimulation of pol delta were blocked by alpha SSB70C, whereas the stimulation of pol alpha by human SSB was unaffected by
this antibody. Conversely, alpha SSB70A, -70B, and -34A inhibited the stimulation of pol alpha, but they had no effect on
DNA unwinding and pol delta stimulation. None of the antibodies inhibited the binding of SSB to single-stranded DNA. These
results suggest that DNA unwinding and stimulation of pol alpha and pol delta are required functions of human SSB in SV40
DNA replication. The human SSB 70-kDa subunit appears to be required for DNA unwinding and pol delta stimulation, whereas
both the 70- and 34-kDa subunits may be involved in the stimulation of pol alpha.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>2159011</pmid><doi>10.1016/S0021-9258(19)39170-7</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 1990-05, Vol.265 (13), p.7693-7700 |
| issn | 0021-9258 1083-351X |
| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection; EZB Electronic Journals Library |
| subjects | Antibodies, Monoclonal Base Sequence Biological and medical sciences Cell Line DNA Probes DNA Replication DNA, Viral - genetics DNA-Binding Proteins - isolation & purification DNA-Binding Proteins - metabolism Fundamental and applied biological sciences. Psychology HeLa Cells - metabolism Humans Kinetics Macromolecular Substances Molecular and cellular biology Molecular genetics Molecular Sequence Data Molecular Weight Oligonucleotide Probes Replication Simian virus 40 - genetics |
| title | The role of human single-stranded DNA binding protein and its individual subunits in simian virus 40 DNA replication |
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