Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus ORF51 is a ChaB homologous gene involved in budded virus production and DNA replication
The baculovirus ChaB proteins are conserved in all completely sequenced Lepidopteran NPVs and are annotated as putative DNA binding proteins. Here we investigated Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) ORF51 ( ha51), one of the ChaB homologues in HearNPV. 5′-RACE rev...
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| Veröffentlicht in: | Virus research 2011, Vol.155 (1), p.203-212 |
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| creator | Zheng, Fangliang Huang, Yi Long, Gang Sun, Xiulian Wang, Hanzhong |
| description | The baculovirus ChaB proteins are conserved in all completely sequenced Lepidopteran NPVs and are annotated as putative DNA binding proteins. Here we investigated
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) ORF51 (
ha51), one of the ChaB homologues in HearNPV. 5′-RACE revealed that
Ha51 is transcribed from a conventional early promoter transcriptional initiator motif (CATT) located at 159nt upstream of ATG. RT-PCR confirmed that
ha51 is an early transcribed gene. To study the function of
Ha51 in the life cycle of HearNPV,
Ha51 knockout and repair bacmids were generated by homologous recombination in
Escherichia coli. Growth curve and DNA replication analyses showed that the levels of budded virus (BV) production and viral DNA accumulation were significantly higher in cells infected with
Ha51 null virus than those infected with wild-type bacmid derived virus. Electron microscopy revealed that polyhedra formation was not affected by the deletion of
Ha51. Bioassay demonstrated that the
Ha51-deleted virus had similar oral infectivity as the wild-type and rescued virus. Western blot analyses suggested that HA51 is a component of the nucleocapsid of BV and occlusion-derived virus as well as the envelope of BV. Immunofluorescence microscopy showed that HA51 protein is mainly localized in the cytoplasm of infected cells. Taken together, our results indicate that, unlike previously characterized baculovirual ChaB genes,
Ha51 is involved in viral DNA replication and BV production and is transcribed in the early stage of infection. |
| doi_str_mv | 10.1016/j.virusres.2010.10.006 |
| format | Article |
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Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) ORF51 (
ha51), one of the ChaB homologues in HearNPV. 5′-RACE revealed that
Ha51 is transcribed from a conventional early promoter transcriptional initiator motif (CATT) located at 159nt upstream of ATG. RT-PCR confirmed that
ha51 is an early transcribed gene. To study the function of
Ha51 in the life cycle of HearNPV,
Ha51 knockout and repair bacmids were generated by homologous recombination in
Escherichia coli. Growth curve and DNA replication analyses showed that the levels of budded virus (BV) production and viral DNA accumulation were significantly higher in cells infected with
Ha51 null virus than those infected with wild-type bacmid derived virus. Electron microscopy revealed that polyhedra formation was not affected by the deletion of
Ha51. Bioassay demonstrated that the
Ha51-deleted virus had similar oral infectivity as the wild-type and rescued virus. Western blot analyses suggested that HA51 is a component of the nucleocapsid of BV and occlusion-derived virus as well as the envelope of BV. Immunofluorescence microscopy showed that HA51 protein is mainly localized in the cytoplasm of infected cells. Taken together, our results indicate that, unlike previously characterized baculovirual ChaB genes,
Ha51 is involved in viral DNA replication and BV production and is transcribed in the early stage of infection.</description><identifier>ISSN: 0168-1702</identifier><identifier>EISSN: 1872-7492</identifier><identifier>DOI: 10.1016/j.virusres.2010.10.006</identifier><identifier>PMID: 20951176</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Animals ; Baculovirus ; bioassays ; BV production ; Cell Line ; ChaB ; cytoplasm ; DNA ; DNA Replication ; DNA-binding proteins ; DNA-Binding Proteins - genetics ; DNA-Binding Proteins - metabolism ; electron microscopy ; Escherichia coli ; Escherichia coli - genetics ; fluorescence microscopy ; Gene Deletion ; Gene Expression Profiling ; Gene Expression Regulation, Viral ; genes ; HearNPV ORF51 ( Ha51) ; Helicoverpa armigera ; Helicoverpa armigera nucleopolyhedrovirus ; homologous recombination ; infection ; Lepidoptera ; Lepidoptera - virology ; Nuclear polyhedrosis virus ; nucleocapsid ; Nucleopolyhedrovirus - physiology ; nucleotide motifs ; pathogenicity ; promoter regions ; reverse transcriptase polymerase chain reaction ; sequence homology ; transcription (genetics) ; Transcription, Genetic ; Viral Proteins - genetics ; Viral Proteins - metabolism ; Virus Release ; Virus Replication ; viruses ; Western blotting</subject><ispartof>Virus research, 2011, Vol.155 (1), p.203-212</ispartof><rights>2010 Elsevier B.V.</rights><rights>Copyright © 2010 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c423t-51bdd8cbbc7b6b144123f501b28ecf8a2c14d1ab3925dd7ac4660714026a07bf3</citedby><cites>FETCH-LOGICAL-c423t-51bdd8cbbc7b6b144123f501b28ecf8a2c14d1ab3925dd7ac4660714026a07bf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.virusres.2010.10.006$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,4024,27923,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20951176$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zheng, Fangliang</creatorcontrib><creatorcontrib>Huang, Yi</creatorcontrib><creatorcontrib>Long, Gang</creatorcontrib><creatorcontrib>Sun, Xiulian</creatorcontrib><creatorcontrib>Wang, Hanzhong</creatorcontrib><title>Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus ORF51 is a ChaB homologous gene involved in budded virus production and DNA replication</title><title>Virus research</title><addtitle>Virus Res</addtitle><description>The baculovirus ChaB proteins are conserved in all completely sequenced Lepidopteran NPVs and are annotated as putative DNA binding proteins. Here we investigated
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) ORF51 (
ha51), one of the ChaB homologues in HearNPV. 5′-RACE revealed that
Ha51 is transcribed from a conventional early promoter transcriptional initiator motif (CATT) located at 159nt upstream of ATG. RT-PCR confirmed that
ha51 is an early transcribed gene. To study the function of
Ha51 in the life cycle of HearNPV,
Ha51 knockout and repair bacmids were generated by homologous recombination in
Escherichia coli. Growth curve and DNA replication analyses showed that the levels of budded virus (BV) production and viral DNA accumulation were significantly higher in cells infected with
Ha51 null virus than those infected with wild-type bacmid derived virus. Electron microscopy revealed that polyhedra formation was not affected by the deletion of
Ha51. Bioassay demonstrated that the
Ha51-deleted virus had similar oral infectivity as the wild-type and rescued virus. Western blot analyses suggested that HA51 is a component of the nucleocapsid of BV and occlusion-derived virus as well as the envelope of BV. Immunofluorescence microscopy showed that HA51 protein is mainly localized in the cytoplasm of infected cells. Taken together, our results indicate that, unlike previously characterized baculovirual ChaB genes,
Ha51 is involved in viral DNA replication and BV production and is transcribed in the early stage of infection.</description><subject>Animals</subject><subject>Baculovirus</subject><subject>bioassays</subject><subject>BV production</subject><subject>Cell Line</subject><subject>ChaB</subject><subject>cytoplasm</subject><subject>DNA</subject><subject>DNA Replication</subject><subject>DNA-binding proteins</subject><subject>DNA-Binding Proteins - genetics</subject><subject>DNA-Binding Proteins - metabolism</subject><subject>electron microscopy</subject><subject>Escherichia coli</subject><subject>Escherichia coli - genetics</subject><subject>fluorescence microscopy</subject><subject>Gene Deletion</subject><subject>Gene Expression Profiling</subject><subject>Gene Expression Regulation, Viral</subject><subject>genes</subject><subject>HearNPV ORF51 ( Ha51)</subject><subject>Helicoverpa armigera</subject><subject>Helicoverpa armigera nucleopolyhedrovirus</subject><subject>homologous recombination</subject><subject>infection</subject><subject>Lepidoptera</subject><subject>Lepidoptera - virology</subject><subject>Nuclear polyhedrosis virus</subject><subject>nucleocapsid</subject><subject>Nucleopolyhedrovirus - physiology</subject><subject>nucleotide motifs</subject><subject>pathogenicity</subject><subject>promoter regions</subject><subject>reverse transcriptase polymerase chain reaction</subject><subject>sequence homology</subject><subject>transcription (genetics)</subject><subject>Transcription, Genetic</subject><subject>Viral Proteins - genetics</subject><subject>Viral Proteins - metabolism</subject><subject>Virus Release</subject><subject>Virus Replication</subject><subject>viruses</subject><subject>Western blotting</subject><issn>0168-1702</issn><issn>1872-7492</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc9u1DAQxi0EosvCKxTfOGWxncRObpSFUqSKSkDPlv9Msl4lcbA3kfogvC9Os-Xak8fj38x84w-hS0p2lFD-8bibXZhigLhj5DG5I4S_QBtaCZaJomYv0SaBVUYFYRfoTYxHkohc8NfogpG6pFTwDfp7A50zfoYwKqxC71oICkc3tB3gYTIdeKPG6Oz5Mvru4QA2-Mfx-O7ndUmxi1jh_UF9xgff-863Pj21MAB2w-y7GWwKsJ6sTdFaOAZvJ3NyfsBqsPjLjyscYExS1JJ7i141qovw7nxu0f3119_7m-z27tv3_dVtZgqWn7KSamsro7URmmtaFJTlTUmoZhWYplLM0MJSpfOaldYKZQrOiaAFYVwRoZt8iz6sfZOcPxPEk-xdNNB1aoC0g6w4ySuai_p5ktGiEqIuE8lX0gQfk0GNHIPrVXiQlMjFO3mUT97JxbslvzizRZfnEZPuwf4vezIrAe9XoFFeqja4KO9_pQ4lIbSiZckS8WklIH3a7CDIaBwMBqwLYE7Sevecin_ckbql</recordid><startdate>2011</startdate><enddate>2011</enddate><creator>Zheng, Fangliang</creator><creator>Huang, Yi</creator><creator>Long, Gang</creator><creator>Sun, Xiulian</creator><creator>Wang, Hanzhong</creator><general>Elsevier B.V</general><scope>FBQ</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>7X8</scope><scope>7SS</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>2011</creationdate><title>Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus ORF51 is a ChaB homologous gene involved in budded virus production and DNA replication</title><author>Zheng, Fangliang ; Huang, Yi ; Long, Gang ; Sun, Xiulian ; Wang, Hanzhong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c423t-51bdd8cbbc7b6b144123f501b28ecf8a2c14d1ab3925dd7ac4660714026a07bf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Baculovirus</topic><topic>bioassays</topic><topic>BV production</topic><topic>Cell Line</topic><topic>ChaB</topic><topic>cytoplasm</topic><topic>DNA</topic><topic>DNA Replication</topic><topic>DNA-binding proteins</topic><topic>DNA-Binding Proteins - genetics</topic><topic>DNA-Binding Proteins - metabolism</topic><topic>electron microscopy</topic><topic>Escherichia coli</topic><topic>Escherichia coli - genetics</topic><topic>fluorescence microscopy</topic><topic>Gene Deletion</topic><topic>Gene Expression Profiling</topic><topic>Gene Expression Regulation, Viral</topic><topic>genes</topic><topic>HearNPV ORF51 ( Ha51)</topic><topic>Helicoverpa armigera</topic><topic>Helicoverpa armigera nucleopolyhedrovirus</topic><topic>homologous recombination</topic><topic>infection</topic><topic>Lepidoptera</topic><topic>Lepidoptera - virology</topic><topic>Nuclear polyhedrosis virus</topic><topic>nucleocapsid</topic><topic>Nucleopolyhedrovirus - physiology</topic><topic>nucleotide motifs</topic><topic>pathogenicity</topic><topic>promoter regions</topic><topic>reverse transcriptase polymerase chain reaction</topic><topic>sequence homology</topic><topic>transcription (genetics)</topic><topic>Transcription, Genetic</topic><topic>Viral Proteins - genetics</topic><topic>Viral Proteins - metabolism</topic><topic>Virus Release</topic><topic>Virus Replication</topic><topic>viruses</topic><topic>Western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zheng, Fangliang</creatorcontrib><creatorcontrib>Huang, Yi</creatorcontrib><creatorcontrib>Long, Gang</creatorcontrib><creatorcontrib>Sun, Xiulian</creatorcontrib><creatorcontrib>Wang, Hanzhong</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Virus research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zheng, Fangliang</au><au>Huang, Yi</au><au>Long, Gang</au><au>Sun, Xiulian</au><au>Wang, Hanzhong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus ORF51 is a ChaB homologous gene involved in budded virus production and DNA replication</atitle><jtitle>Virus research</jtitle><addtitle>Virus Res</addtitle><date>2011</date><risdate>2011</risdate><volume>155</volume><issue>1</issue><spage>203</spage><epage>212</epage><pages>203-212</pages><issn>0168-1702</issn><eissn>1872-7492</eissn><abstract>The baculovirus ChaB proteins are conserved in all completely sequenced Lepidopteran NPVs and are annotated as putative DNA binding proteins. Here we investigated
Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HearNPV) ORF51 (
ha51), one of the ChaB homologues in HearNPV. 5′-RACE revealed that
Ha51 is transcribed from a conventional early promoter transcriptional initiator motif (CATT) located at 159nt upstream of ATG. RT-PCR confirmed that
ha51 is an early transcribed gene. To study the function of
Ha51 in the life cycle of HearNPV,
Ha51 knockout and repair bacmids were generated by homologous recombination in
Escherichia coli. Growth curve and DNA replication analyses showed that the levels of budded virus (BV) production and viral DNA accumulation were significantly higher in cells infected with
Ha51 null virus than those infected with wild-type bacmid derived virus. Electron microscopy revealed that polyhedra formation was not affected by the deletion of
Ha51. Bioassay demonstrated that the
Ha51-deleted virus had similar oral infectivity as the wild-type and rescued virus. Western blot analyses suggested that HA51 is a component of the nucleocapsid of BV and occlusion-derived virus as well as the envelope of BV. Immunofluorescence microscopy showed that HA51 protein is mainly localized in the cytoplasm of infected cells. Taken together, our results indicate that, unlike previously characterized baculovirual ChaB genes,
Ha51 is involved in viral DNA replication and BV production and is transcribed in the early stage of infection.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>20951176</pmid><doi>10.1016/j.virusres.2010.10.006</doi><tpages>10</tpages></addata></record> |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Animals Baculovirus bioassays BV production Cell Line ChaB cytoplasm DNA DNA Replication DNA-binding proteins DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism electron microscopy Escherichia coli Escherichia coli - genetics fluorescence microscopy Gene Deletion Gene Expression Profiling Gene Expression Regulation, Viral genes HearNPV ORF51 ( Ha51) Helicoverpa armigera Helicoverpa armigera nucleopolyhedrovirus homologous recombination infection Lepidoptera Lepidoptera - virology Nuclear polyhedrosis virus nucleocapsid Nucleopolyhedrovirus - physiology nucleotide motifs pathogenicity promoter regions reverse transcriptase polymerase chain reaction sequence homology transcription (genetics) Transcription, Genetic Viral Proteins - genetics Viral Proteins - metabolism Virus Release Virus Replication viruses Western blotting |
| title | Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus ORF51 is a ChaB homologous gene involved in budded virus production and DNA replication |
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