Role of conserved residues in the activity of adenovirus preterminal protein
Centre for Biomolecular Sciences, School of Biology, University of St Andrews, Biomolecular Sciences Building, North Haugh, St Andrews, Fife KY16 9ST, UK 1 Author for correspondence: Ronald Hay. Fax +44 1334 462595. e-mail rth{at}st-andrews.ac.uk Preterminal protein (pTP) is a component of the prein...
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Veröffentlicht in: | Journal of general virology 2001-08, Vol.82 (8), p.1917-1927 |
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container_end_page | 1927 |
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container_issue | 8 |
container_start_page | 1917 |
container_title | Journal of general virology |
container_volume | 82 |
creator | Botting, Catherine H Hay, Ronald T |
description | Centre for Biomolecular Sciences, School of Biology, University of St Andrews, Biomolecular Sciences Building, North Haugh, St Andrews, Fife KY16 9ST, UK 1
Author for correspondence: Ronald Hay. Fax +44 1334 462595. e-mail rth{at}st-andrews.ac.uk
Preterminal protein (pTP) is a component of the preinitiation complex which forms at the adenovirus origin of DNA replication and acts as the protein primer during DNA synthesis. In order to determine the role of various regions of the molecule a series of 18 mutations was introduced into conserved motifs of pTP which were predicted to be surface exposed, and the mutants expressed in insect cells using a baculovirus expression system. Their ability to initiate DNA replication was assessed and the effect the mutations have on the individual interactions which contribute to the formation of the pre-initiation complex was determined. Classes of mutants could be identified which were unable to bind DNA or interact with the adenovirus DNA polymerase, but one class of mutants retained these activities and yet failed to initiate DNA replication. These mutants therefore identify regions of pTP required for different aspects of adenovirus DNA replication. |
doi_str_mv | 10.1099/0022-1317-82-8-1917 |
format | Article |
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Author for correspondence: Ronald Hay. Fax +44 1334 462595. e-mail rth{at}st-andrews.ac.uk
Preterminal protein (pTP) is a component of the preinitiation complex which forms at the adenovirus origin of DNA replication and acts as the protein primer during DNA synthesis. In order to determine the role of various regions of the molecule a series of 18 mutations was introduced into conserved motifs of pTP which were predicted to be surface exposed, and the mutants expressed in insect cells using a baculovirus expression system. Their ability to initiate DNA replication was assessed and the effect the mutations have on the individual interactions which contribute to the formation of the pre-initiation complex was determined. Classes of mutants could be identified which were unable to bind DNA or interact with the adenovirus DNA polymerase, but one class of mutants retained these activities and yet failed to initiate DNA replication. These mutants therefore identify regions of pTP required for different aspects of adenovirus DNA replication.</description><identifier>ISSN: 0022-1317</identifier><identifier>EISSN: 1465-2099</identifier><identifier>DOI: 10.1099/0022-1317-82-8-1917</identifier><identifier>PMID: 11457998</identifier><language>eng</language><publisher>England: Soc General Microbiol</publisher><subject>Adenovirus ; Adenoviruses, Human ; Amino Acid Sequence ; Animals ; Baculoviridae - genetics ; Baculovirus ; Cell Line ; DNA Replication ; DNA, Single-Stranded ; DNA, Viral - biosynthesis ; Genetic Vectors ; Humans ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Phosphoproteins - genetics ; Phosphoproteins - physiology ; Protein Binding ; Protein Precursors - genetics ; Protein Precursors - physiology ; terminal protein ; Viral Proteins ; Virus Replication</subject><ispartof>Journal of general virology, 2001-08, Vol.82 (8), p.1917-1927</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c410t-bafc033606edf4d568091eb6b7b43b72e1736b2e5ee1727b274602d1b42dbbcc3</citedby><cites>FETCH-LOGICAL-c410t-bafc033606edf4d568091eb6b7b43b72e1736b2e5ee1727b274602d1b42dbbcc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,3733,3734,27905,27906</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11457998$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Botting, Catherine H</creatorcontrib><creatorcontrib>Hay, Ronald T</creatorcontrib><title>Role of conserved residues in the activity of adenovirus preterminal protein</title><title>Journal of general virology</title><addtitle>J Gen Virol</addtitle><description>Centre for Biomolecular Sciences, School of Biology, University of St Andrews, Biomolecular Sciences Building, North Haugh, St Andrews, Fife KY16 9ST, UK 1
Author for correspondence: Ronald Hay. Fax +44 1334 462595. e-mail rth{at}st-andrews.ac.uk
Preterminal protein (pTP) is a component of the preinitiation complex which forms at the adenovirus origin of DNA replication and acts as the protein primer during DNA synthesis. In order to determine the role of various regions of the molecule a series of 18 mutations was introduced into conserved motifs of pTP which were predicted to be surface exposed, and the mutants expressed in insect cells using a baculovirus expression system. Their ability to initiate DNA replication was assessed and the effect the mutations have on the individual interactions which contribute to the formation of the pre-initiation complex was determined. Classes of mutants could be identified which were unable to bind DNA or interact with the adenovirus DNA polymerase, but one class of mutants retained these activities and yet failed to initiate DNA replication. These mutants therefore identify regions of pTP required for different aspects of adenovirus DNA replication.</description><subject>Adenovirus</subject><subject>Adenoviruses, Human</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Baculoviridae - genetics</subject><subject>Baculovirus</subject><subject>Cell Line</subject><subject>DNA Replication</subject><subject>DNA, Single-Stranded</subject><subject>DNA, Viral - biosynthesis</subject><subject>Genetic Vectors</subject><subject>Humans</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>Phosphoproteins - genetics</subject><subject>Phosphoproteins - physiology</subject><subject>Protein Binding</subject><subject>Protein Precursors - genetics</subject><subject>Protein Precursors - physiology</subject><subject>terminal protein</subject><subject>Viral Proteins</subject><subject>Virus Replication</subject><issn>0022-1317</issn><issn>1465-2099</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkN9LwzAQx4Mobk7_AkH6JL5EkzRtmkcZ_oKBIPocmuS6RdpmJt1k_70pG-7pDu5z3zs-CF1Tck-JlA-EMIZpTgWuGK4wlVScoCnlZYFZmp-i6T8xQRcxfhNCOS_EOZpQmqqU1RQtPnwLmW8y4_sIYQs2CxCd3UDMXJ8NK8hqM7itG3YjVVvo_daFTczWAQYInevrNvV-ANdforOmbiNcHeoMfT0_fc5f8eL95W3-uMCGUzJgXTeG5HlJSrANt0VZEUlBl1ponmvBgIq81AwKSB0TmgleEmap5sxqbUw-Q7f73HT3J306qM5FA21b9-A3UVEhmSyYTGC-B03wMQZo1Dq4rg47RYkaJapRkRoVqYqpSo0S09bNIX6jO7DHnYO1BNztgZVbrn5dALWEvnPpiHZeJT3HrD_Mtnvl</recordid><startdate>20010801</startdate><enddate>20010801</enddate><creator>Botting, Catherine H</creator><creator>Hay, Ronald T</creator><general>Soc General Microbiol</general><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></search><sort><creationdate>20010801</creationdate><title>Role of conserved residues in the activity of adenovirus preterminal protein</title><author>Botting, Catherine H ; Hay, Ronald T</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c410t-bafc033606edf4d568091eb6b7b43b72e1736b2e5ee1727b274602d1b42dbbcc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Adenovirus</topic><topic>Adenoviruses, Human</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Baculoviridae - genetics</topic><topic>Baculovirus</topic><topic>Cell Line</topic><topic>DNA Replication</topic><topic>DNA, Single-Stranded</topic><topic>DNA, Viral - biosynthesis</topic><topic>Genetic Vectors</topic><topic>Humans</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>Phosphoproteins - genetics</topic><topic>Phosphoproteins - physiology</topic><topic>Protein Binding</topic><topic>Protein Precursors - genetics</topic><topic>Protein Precursors - physiology</topic><topic>terminal protein</topic><topic>Viral Proteins</topic><topic>Virus Replication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Botting, Catherine H</creatorcontrib><creatorcontrib>Hay, Ronald T</creatorcontrib><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><jtitle>Journal of general virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Botting, Catherine H</au><au>Hay, Ronald T</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Role of conserved residues in the activity of adenovirus preterminal protein</atitle><jtitle>Journal of general virology</jtitle><addtitle>J Gen Virol</addtitle><date>2001-08-01</date><risdate>2001</risdate><volume>82</volume><issue>8</issue><spage>1917</spage><epage>1927</epage><pages>1917-1927</pages><issn>0022-1317</issn><eissn>1465-2099</eissn><abstract>Centre for Biomolecular Sciences, School of Biology, University of St Andrews, Biomolecular Sciences Building, North Haugh, St Andrews, Fife KY16 9ST, UK 1
Author for correspondence: Ronald Hay. Fax +44 1334 462595. e-mail rth{at}st-andrews.ac.uk
Preterminal protein (pTP) is a component of the preinitiation complex which forms at the adenovirus origin of DNA replication and acts as the protein primer during DNA synthesis. In order to determine the role of various regions of the molecule a series of 18 mutations was introduced into conserved motifs of pTP which were predicted to be surface exposed, and the mutants expressed in insect cells using a baculovirus expression system. Their ability to initiate DNA replication was assessed and the effect the mutations have on the individual interactions which contribute to the formation of the pre-initiation complex was determined. Classes of mutants could be identified which were unable to bind DNA or interact with the adenovirus DNA polymerase, but one class of mutants retained these activities and yet failed to initiate DNA replication. These mutants therefore identify regions of pTP required for different aspects of adenovirus DNA replication.</abstract><cop>England</cop><pub>Soc General Microbiol</pub><pmid>11457998</pmid><doi>10.1099/0022-1317-82-8-1917</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Adenovirus Adenoviruses, Human Amino Acid Sequence Animals Baculoviridae - genetics Baculovirus Cell Line DNA Replication DNA, Single-Stranded DNA, Viral - biosynthesis Genetic Vectors Humans Molecular Sequence Data Mutagenesis, Site-Directed Phosphoproteins - genetics Phosphoproteins - physiology Protein Binding Protein Precursors - genetics Protein Precursors - physiology terminal protein Viral Proteins Virus Replication |
title | Role of conserved residues in the activity of adenovirus preterminal protein |
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