Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3
We have identified both biochemically and genetically a protein domain within the simian virus 40 virion protein Vp3, and within Vp2 since its carboxyl two-thirds are identical to the full-length Vp3, that binds DNA in a sequence nonspecific manner. Both the Vp2 and Vp3 (Vp2/3) components of SV40 an...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 1993-10, Vol.268 (28), p.20877-20883 |
|---|---|
| Hauptverfasser: | , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 20883 |
|---|---|
| container_issue | 28 |
| container_start_page | 20877 |
| container_title | The Journal of biological chemistry |
| container_volume | 268 |
| creator | CLEVER, J DEAN, D. A KASAMATSU, H |
| description | We have identified both biochemically and genetically a protein domain within the simian virus 40 virion protein Vp3, and
within Vp2 since its carboxyl two-thirds are identical to the full-length Vp3, that binds DNA in a sequence nonspecific manner.
Both the Vp2 and Vp3 (Vp2/3) components of SV40 and mutant SV40(202T) bound either SV40 or pBR322 DNA equally well. Wild type
and mutant Vp2/3 proteins, expressed as fusion proteins with glutathione S-transferase (GST), were tested for their ability
to bind DNA. GST-Vp3 bound DNA at physiological salt concentrations with an apparent Kd of 2.5 x 10(-8) M and also bound RNA
with 4-fold higher affinity. Over 90% of the nucleic acid binding, and all of the activity, was lost upon removal of the carboxyl-terminal
13 and 35 residues, respectively. The DNA binding domain was shown to be distinct and separable from the Vp2/3 nuclear transport
signal since mutations within the nuclear transport signal that reduce or abolish nuclear localization of Vp2/3 had no effect
on the DNA binding activity of mutant Vp2/3 fusion proteins. The carboxyl-terminal 40 residues of Vp2/3 in the form of a beta-galactosidase
fusion protein, F6, are sufficient for DNA binding and may cause compaction of the DNA. The significance of this DNA binding
and possible compaction are discussed in relation to the assembly of virion particles. |
| doi_str_mv | 10.1016/s0021-9258(19)36868-1 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_75986667</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>16653019</sourcerecordid><originalsourceid>FETCH-LOGICAL-c506t-996117df0ff08201688cf0a9372f4a1e6645e36982341067b5fdb73e98922a1e3</originalsourceid><addsrcrecordid>eNqFkE1v1DAQhi0EKtvCT6jkA0JwCMzYiT-OVQulUgUHKOJmOY7dHbRxljgL4t_jpavlyGikObzPfL2MnSO8QUD1tgAIbKzozCu0r6UyyjT4iK0QjGxkh98es9URecpOS_kONVqLJ-zEtKCtgBW7uxliXihR8AtNmU-Je3718YL3lAfK93yYRk-Z1yw0ks_8J827wlvgwW8LDXw7T0ukXPjXreA-D7XKZ-xJ8psSnx_qGbt7_-7L5Yfm9tP1zeXFbRM6UEtjrULUQ4KUwIj6lDEhgbdSi9R6jEq1XZTKGiFbBKX7Lg29ltEaK0TV5Rl7-TC3HvFjF8viRiohbjY-x2lXnO6sUUrp_4KoVCcBbQW7BzDMUylzTG470-jn3w7B7X13n_emur2pDq3767vD2nd-WLDrxzgcuw5GV_3FQfcl-E2afQ5UjpisL9rW_MPWdL_-RXN0PU1hHUcn6hpRE4zW8g8Lu5OX</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16653019</pqid></control><display><type>article</type><title>Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3</title><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Alma/SFX Local Collection</source><creator>CLEVER, J ; DEAN, D. A ; KASAMATSU, H</creator><creatorcontrib>CLEVER, J ; DEAN, D. A ; KASAMATSU, H</creatorcontrib><description>We have identified both biochemically and genetically a protein domain within the simian virus 40 virion protein Vp3, and
within Vp2 since its carboxyl two-thirds are identical to the full-length Vp3, that binds DNA in a sequence nonspecific manner.
Both the Vp2 and Vp3 (Vp2/3) components of SV40 and mutant SV40(202T) bound either SV40 or pBR322 DNA equally well. Wild type
and mutant Vp2/3 proteins, expressed as fusion proteins with glutathione S-transferase (GST), were tested for their ability
to bind DNA. GST-Vp3 bound DNA at physiological salt concentrations with an apparent Kd of 2.5 x 10(-8) M and also bound RNA
with 4-fold higher affinity. Over 90% of the nucleic acid binding, and all of the activity, was lost upon removal of the carboxyl-terminal
13 and 35 residues, respectively. The DNA binding domain was shown to be distinct and separable from the Vp2/3 nuclear transport
signal since mutations within the nuclear transport signal that reduce or abolish nuclear localization of Vp2/3 had no effect
on the DNA binding activity of mutant Vp2/3 fusion proteins. The carboxyl-terminal 40 residues of Vp2/3 in the form of a beta-galactosidase
fusion protein, F6, are sufficient for DNA binding and may cause compaction of the DNA. The significance of this DNA binding
and possible compaction are discussed in relation to the assembly of virion particles.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/s0021-9258(19)36868-1</identifier><identifier>PMID: 8407920</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Biochemistry and Molecular Biology</publisher><subject>Animals ; beta-Galactosidase - metabolism ; Binding Sites ; Biological and medical sciences ; Blotting, Southern ; Blotting, Western ; Capsid - metabolism ; Capsid Proteins ; Cell Nucleus - metabolism ; Cells, Cultured ; Chlorocebus aethiops ; DNA, Recombinant - metabolism ; DNA, Viral - metabolism ; Fundamental and applied biological sciences. Psychology ; Glutathione Transferase - metabolism ; Microbiology ; Recombinant Fusion Proteins - metabolism ; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains ; Simian virus 40 - genetics ; Simian virus 40 - metabolism ; Solutions ; Virion - metabolism ; Virology</subject><ispartof>The Journal of biological chemistry, 1993-10, Vol.268 (28), p.20877-20883</ispartof><rights>1994 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c506t-996117df0ff08201688cf0a9372f4a1e6645e36982341067b5fdb73e98922a1e3</citedby><cites>FETCH-LOGICAL-c506t-996117df0ff08201688cf0a9372f4a1e6645e36982341067b5fdb73e98922a1e3</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=3823948$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8407920$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>CLEVER, J</creatorcontrib><creatorcontrib>DEAN, D. A</creatorcontrib><creatorcontrib>KASAMATSU, H</creatorcontrib><title>Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>We have identified both biochemically and genetically a protein domain within the simian virus 40 virion protein Vp3, and
within Vp2 since its carboxyl two-thirds are identical to the full-length Vp3, that binds DNA in a sequence nonspecific manner.
Both the Vp2 and Vp3 (Vp2/3) components of SV40 and mutant SV40(202T) bound either SV40 or pBR322 DNA equally well. Wild type
and mutant Vp2/3 proteins, expressed as fusion proteins with glutathione S-transferase (GST), were tested for their ability
to bind DNA. GST-Vp3 bound DNA at physiological salt concentrations with an apparent Kd of 2.5 x 10(-8) M and also bound RNA
with 4-fold higher affinity. Over 90% of the nucleic acid binding, and all of the activity, was lost upon removal of the carboxyl-terminal
13 and 35 residues, respectively. The DNA binding domain was shown to be distinct and separable from the Vp2/3 nuclear transport
signal since mutations within the nuclear transport signal that reduce or abolish nuclear localization of Vp2/3 had no effect
on the DNA binding activity of mutant Vp2/3 fusion proteins. The carboxyl-terminal 40 residues of Vp2/3 in the form of a beta-galactosidase
fusion protein, F6, are sufficient for DNA binding and may cause compaction of the DNA. The significance of this DNA binding
and possible compaction are discussed in relation to the assembly of virion particles.</description><subject>Animals</subject><subject>beta-Galactosidase - metabolism</subject><subject>Binding Sites</subject><subject>Biological and medical sciences</subject><subject>Blotting, Southern</subject><subject>Blotting, Western</subject><subject>Capsid - metabolism</subject><subject>Capsid Proteins</subject><subject>Cell Nucleus - metabolism</subject><subject>Cells, Cultured</subject><subject>Chlorocebus aethiops</subject><subject>DNA, Recombinant - metabolism</subject><subject>DNA, Viral - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glutathione Transferase - metabolism</subject><subject>Microbiology</subject><subject>Recombinant Fusion Proteins - metabolism</subject><subject>Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains</subject><subject>Simian virus 40 - genetics</subject><subject>Simian virus 40 - metabolism</subject><subject>Solutions</subject><subject>Virion - metabolism</subject><subject>Virology</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1v1DAQhi0EKtvCT6jkA0JwCMzYiT-OVQulUgUHKOJmOY7dHbRxljgL4t_jpavlyGikObzPfL2MnSO8QUD1tgAIbKzozCu0r6UyyjT4iK0QjGxkh98es9URecpOS_kONVqLJ-zEtKCtgBW7uxliXihR8AtNmU-Je3718YL3lAfK93yYRk-Z1yw0ks_8J827wlvgwW8LDXw7T0ukXPjXreA-D7XKZ-xJ8psSnx_qGbt7_-7L5Yfm9tP1zeXFbRM6UEtjrULUQ4KUwIj6lDEhgbdSi9R6jEq1XZTKGiFbBKX7Lg29ltEaK0TV5Rl7-TC3HvFjF8viRiohbjY-x2lXnO6sUUrp_4KoVCcBbQW7BzDMUylzTG470-jn3w7B7X13n_emur2pDq3767vD2nd-WLDrxzgcuw5GV_3FQfcl-E2afQ5UjpisL9rW_MPWdL_-RXN0PU1hHUcn6hpRE4zW8g8Lu5OX</recordid><startdate>19931005</startdate><enddate>19931005</enddate><creator>CLEVER, J</creator><creator>DEAN, D. A</creator><creator>KASAMATSU, H</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>19931005</creationdate><title>Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3</title><author>CLEVER, J ; DEAN, D. A ; KASAMATSU, H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c506t-996117df0ff08201688cf0a9372f4a1e6645e36982341067b5fdb73e98922a1e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Animals</topic><topic>beta-Galactosidase - metabolism</topic><topic>Binding Sites</topic><topic>Biological and medical sciences</topic><topic>Blotting, Southern</topic><topic>Blotting, Western</topic><topic>Capsid - metabolism</topic><topic>Capsid Proteins</topic><topic>Cell Nucleus - metabolism</topic><topic>Cells, Cultured</topic><topic>Chlorocebus aethiops</topic><topic>DNA, Recombinant - metabolism</topic><topic>DNA, Viral - metabolism</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Glutathione Transferase - metabolism</topic><topic>Microbiology</topic><topic>Recombinant Fusion Proteins - metabolism</topic><topic>Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains</topic><topic>Simian virus 40 - genetics</topic><topic>Simian virus 40 - metabolism</topic><topic>Solutions</topic><topic>Virion - metabolism</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>CLEVER, J</creatorcontrib><creatorcontrib>DEAN, D. A</creatorcontrib><creatorcontrib>KASAMATSU, H</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>CLEVER, J</au><au>DEAN, D. A</au><au>KASAMATSU, H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1993-10-05</date><risdate>1993</risdate><volume>268</volume><issue>28</issue><spage>20877</spage><epage>20883</epage><pages>20877-20883</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>We have identified both biochemically and genetically a protein domain within the simian virus 40 virion protein Vp3, and
within Vp2 since its carboxyl two-thirds are identical to the full-length Vp3, that binds DNA in a sequence nonspecific manner.
Both the Vp2 and Vp3 (Vp2/3) components of SV40 and mutant SV40(202T) bound either SV40 or pBR322 DNA equally well. Wild type
and mutant Vp2/3 proteins, expressed as fusion proteins with glutathione S-transferase (GST), were tested for their ability
to bind DNA. GST-Vp3 bound DNA at physiological salt concentrations with an apparent Kd of 2.5 x 10(-8) M and also bound RNA
with 4-fold higher affinity. Over 90% of the nucleic acid binding, and all of the activity, was lost upon removal of the carboxyl-terminal
13 and 35 residues, respectively. The DNA binding domain was shown to be distinct and separable from the Vp2/3 nuclear transport
signal since mutations within the nuclear transport signal that reduce or abolish nuclear localization of Vp2/3 had no effect
on the DNA binding activity of mutant Vp2/3 fusion proteins. The carboxyl-terminal 40 residues of Vp2/3 in the form of a beta-galactosidase
fusion protein, F6, are sufficient for DNA binding and may cause compaction of the DNA. The significance of this DNA binding
and possible compaction are discussed in relation to the assembly of virion particles.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>8407920</pmid><doi>10.1016/s0021-9258(19)36868-1</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 1993-10, Vol.268 (28), p.20877-20883 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_75986667 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Animals beta-Galactosidase - metabolism Binding Sites Biological and medical sciences Blotting, Southern Blotting, Western Capsid - metabolism Capsid Proteins Cell Nucleus - metabolism Cells, Cultured Chlorocebus aethiops DNA, Recombinant - metabolism DNA, Viral - metabolism Fundamental and applied biological sciences. Psychology Glutathione Transferase - metabolism Microbiology Recombinant Fusion Proteins - metabolism Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains Simian virus 40 - genetics Simian virus 40 - metabolism Solutions Virion - metabolism Virology |
| title | Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3 |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-13T23%3A30%3A33IST&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=Identification%20of%20a%20DNA%20binding%20domain%20in%20simian%20virus%2040%20capsid%20proteins%20Vp2%20and%20Vp3&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=CLEVER,%20J&rft.date=1993-10-05&rft.volume=268&rft.issue=28&rft.spage=20877&rft.epage=20883&rft.pages=20877-20883&rft.issn=0021-9258&rft.eissn=1083-351X&rft.coden=JBCHA3&rft_id=info:doi/10.1016/s0021-9258(19)36868-1&rft_dat=%3Cproquest_cross%3E16653019%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=16653019&rft_id=info:pmid/8407920&rfr_iscdi=true |