Adenovirus terminal protein mediates both nuclear matrix association and efficient transcription of adenovirus DNA.
Adenovirus DNA is tightly bound to the nuclear matrix throughout the course of infection. Analysis of adenovirus DNA from infected HeLa cell nuclei after extraction with lithium diiodosalicylate and digestion with restriction enzymes demonstrated that the sites of tightest attachment occur in the terminal fragments of the linear viral chromosome. Analysis of viruses mutated in the precursor terminal protein coding sequence demonstrated that the terminal protein, which is covalently attached to the 5' end of each DNA strand, mediates the tight binding. Virions containing chromosomes with mutant terminal proteins were unpackaged and viral DNA accumulated in the nucleus at a normal rate and competed for the limiting component during transcription complex formation, but their early genes were transcribed at reduced efficiency by both RNA polymerases II and III. The transcriptional defects were not complemented by coinfection with a wild-type virus. We propose that the adenovirus chromosome may exist as a single chromatin domain during infection and that binding of DNA to the nuclear matrix may play a critical role in adenovirus transcription.
Schaack
J
J
Howard Hughes Medical Institute, Department of Biology, Princeton University, New Jersey 08544.
Ho
W Y
WY
Freimuth
P
P
Shenk
T
T
D000260:Adenoviruses, Human / Q000235:genetics* / Q000502:physiology; D002843:Chromatin / Q000378:metabolism / Q000648:ultrastructure; D004279:DNA, Viral / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D015530:Nuclear Matrix / Q000378:metabolism*; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000378:metabolism*
D002843:Chromatin; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; C056461:terminal protein, adenovirus