title: Adenovirus protein VII condenses DNA, represses transcription, and associates with transcriptional activator E1A.
abstract: Adenovirus protein VII is the major protein component of the viral nucleoprotein core. It is highly basic, and an estimated 1070 copies associate with each viral genome, forming a tightly condensed DNA-protein complex. We have investigated DNA condensation, transcriptional repression, and specific protein binding by protein VII. Xenopus oocytes were microinjected with mRNA encoding HA-tagged protein VII and prepared for visualization of lampbrush chromosomes. Immunostaining revealed that protein VII associated in a uniform manner across entire chromosomes. Furthermore, the chromosomes were significantly condensed and transcriptionally silenced, as judged by the dramatic disappearance of transcription loops characteristic of lampbrush chromosomes. During infection, the protein VII-DNA complex may be the initial substrate for transcriptional activation by cellular factors and the viral E1A protein. To investigate this possibility, mRNAs encoding E1A and protein VII were comicroinjected into Xenopus oocytes. Interestingly, whereas E1A did not associate with chromosomes in the absence of protein VII, expression of both proteins together resulted in significant association of E1A with lampbrush chromosomes. Binding studies with proteins produced in bacteria or human cells or by in vitro translation showed that E1A and protein VII can interact in vitro. Structure-function analysis revealed that an N-terminal region of E1A is responsible for binding to protein VII. These studies define the in vivo functions of protein VII in DNA binding, condensation, and transcriptional repression and indicate a role in E1A-mediated transcriptional activation of viral genes.
authors.affiliation: Department of Microbiology, University of Virginia Health System, P.O. Box 800734, Charlottesville, VA 22908, USA.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Johnson
authors.forename: Yvonne N
authors.initials: YN
authors.lastname: Osheim
authors.forename: Yuming
authors.initials: Y
authors.lastname: Xue
authors.forename: Margaux R
authors.initials: MR
authors.lastname: Emanuel
authors.forename: Peter W
authors.initials: PW
authors.lastname: Lewis
authors.forename: Alex
authors.initials: A
authors.lastname: Bankovich
authors.forename: Ann L
authors.initials: AL
authors.lastname: Beyer
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Engel
mesh_terms: D017500:Adenovirus E1A Proteins / Q000378:metabolism*; D000260:Adenoviruses, Human / Q000378:metabolism / Q000472:pathogenicity; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002843:Chromatin; D004279:DNA, Viral / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D006801:Humans; D009865:Oocytes / Q000378:metabolism; D011233:Precipitin Tests; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014758:Viral Core Proteins / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014981:Xenopus / Q000254:growth & development
chemical_list: D017500:Adenovirus E1A Proteins; D002843:Chromatin; D004279:DNA, Viral; D014758:Viral Core Proteins; D014764:Viral Proteins