Adenovirus type 5 DNA-protein complexes from formaldehyde cross-linked cells early after infection.
Journal
  Virology.
Citation
  Virology. 312(1):204-12
Publication date
  2003 Jul 20
Authors
  Spector DJ
Johnson JS
Baird NL
Engel DA
Investigators
  David J. Spector
Grant agencies
  National Cancer Institute
National Institute of General Medical Sciences
Grants
  NCI CA60675
NIGMS GM58214
MeSH headings
  Adenoviridae
DNA, Viral
Formaldehyde
Viral Proteins
MeSH qualifiers
  genetics
metabolism
chemistry
Abstract
  We report here the properties of viral DNA-protein complexes that purify with cellular chromatin following formaldehyde cross-linking of intact cells early after infection. The cross-linked viral DNA fractionated into shear-sensitive (S) and shear- resistant (R) components that were separable by sedimentation, which allowed independent characterization. The R component had the density and sedimentation properties expected for DNA-protein complexes and contained intact viral DNA. It accounted for about 50% of the viral DNA recovered at 1.5 h after infection but less than 20% by 4.5 h. The proportion of R component was independent of multiplicity of infection, even at less than one particle per cell. Viral hexon and protein VII, but not protein VI, were detected in the fractions containing the R component. These properties are consistent with those of partially uncoated virions associated with the nuclear envelope. A substantial proportion of the S component viral DNA had the same density as cellular chromatin. Protein VII was the most abundant viral protein present in gradient fractions that contained the S component. Complexes containing USF transcription factor cross-linked to the adenovirus major late promoter were detected by viral chromatin immunoprecipitation of the fractions containing S component. The S component probably contained uncoated nuclear viral DNA that assembles into early viral transcription complexes.
Medline ID
  22773634