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Human cytomegalovirus pUL79 is an elongation factor of RNA polymerase II for viral gene transcription.
Perng, Yi-Chieh; Campbell, Jessica A; Lenschow, Deborah J; Yu, Dong.
Afiliación
  • Perng YC; Department of Molecular Microbiology, Washington University School of Medicine, Saint Louis, Missouri, United States of America.
  • Campbell JA; Department of Medicine, Department of Pathology and Immunology, Washington University School of Medicine, Saint Louis, Missouri, United States of America.
  • Lenschow DJ; Department of Medicine, Department of Pathology and Immunology, Washington University School of Medicine, Saint Louis, Missouri, United States of America.
  • Yu D; Department of Molecular Microbiology, Washington University School of Medicine, Saint Louis, Missouri, United States of America.
PLoS Pathog ; 10(8): e1004350, 2014 Aug.
Article en En | MEDLINE | ID: mdl-25166009
In this study, we have identified a unique mechanism in which human cytomegalovirus (HCMV) protein pUL79 acts as an elongation factor to direct cellular RNA polymerase II for viral transcription during late times of infection. We and others previously reported that pUL79 and its homologues are required for viral transcript accumulation after viral DNA synthesis. We hypothesized that pUL79 represented a unique mechanism to regulate viral transcription at late times during HCMV infection. To test this hypothesis, we analyzed the proteome associated with pUL79 during virus infection by mass spectrometry. We identified both cellular transcriptional factors, including multiple RNA polymerase II (RNAP II) subunits, and novel viral transactivators, including pUL87 and pUL95, as protein binding partners of pUL79. Co-immunoprecipitation (co-IP) followed by immunoblot analysis confirmed the pUL79-RNAP II interaction, and this interaction was independent of any other viral proteins. Using a recombinant HCMV virus where pUL79 protein is conditionally regulated by a protein destabilization domain ddFKBP, we showed that this interaction did not alter the total levels of RNAP II or its recruitment to viral late promoters. Furthermore, pUL79 did not alter the phosphorylation profiles of the RNAP II C-terminal domain, which was critical for transcriptional regulation. Rather, a nuclear run-on assay indicated that, in the absence of pUL79, RNAP II failed to elongate and stalled on the viral DNA. pUL79-dependent RNAP II elongation was required for transcription from all three kinetic classes of viral genes (i.e. immediate-early, early, and late) at late times during virus infection. In contrast, host gene transcription during HCMV infection was independent of pUL79. In summary, we have identified a novel viral mechanism by which pUL79, and potentially other viral factors, regulates the rate of RNAP II transcription machinery on viral transcription during late stages of HCMV infection.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Virales / ARN Polimerasa II / Regulación Viral de la Expresión Génica / Factores de Elongación de Péptidos / Infecciones por Citomegalovirus Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: PLoS Pathog Año: 2014 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Virales / ARN Polimerasa II / Regulación Viral de la Expresión Génica / Factores de Elongación de Péptidos / Infecciones por Citomegalovirus Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: PLoS Pathog Año: 2014 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos