The structure of adenovirion chromatin revealed by ultraviolet light-induced cross-linking

doi:10.1016/0006-291X(81)91591-6

Biochemical and Biophysical Research Communications

Volume 101, Issue 4, 31 August 1981, Pages 1318-1323

https://doi.org/10.1016/0006-291X(81)91591-6 Get rights and content

Abstract

Upon irradiating adenovirion by ultraviolet light, viral DNA and a part of viral protein cosedimented faster than free DNA when examined in the presence of 4 M guanidine HC1 in sucrose gradient, indicating that protein became tightly bound to DNA. By sodium dodecylsulfate polyacrylamide gel electrophoresis, it was verified that the protein is cross-linked covalently to DNA and that polypeptide VII is the only polypeptide species present. Based on the available data on the size of nuclease digested fragments, a model of adenovirion chromatin consisting of a nucleosome-like subunit of polypeptide VII-hexamer was constructed.

References (15)

E. Everitt et al.
Virology
(1975)
M. Brown et al.
Virol
(1980)
M. Green et al.
Virology
(1963)
M.T. Sung et al.
J. Biol. Chem
(1977)
L. Lica et al.
J. Mol. Biol
(1977)
D.T. Brown et al.
J. Virol
(1975)
K. Hosokawa et al.
J. Virol
(1976)

There are more references available in the full text version of this article.

Cited by (8)

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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.
In vitro transcription of a chromatin-like complex of major core protein VII and DNA of adenovirus serotype 2
1986, Biochemical and Biophysical Research Communications
Major core protein VII of adenovirus serotype 2 (Ad2) is thought to play a role as a histone octamer in eukaryotic cells. We compared the template activity of the VII-DNA complex formed $in$ $vitro$ with that of protein-free DNA. Hybridization assay of $in$ $vitro$ transcripts showed that transcription from regions located in the middle of Ad2 DNA decreased when Ad2 DNA formed a complex with VII. This suggests that the chromatin structure plays a role in regulation of transcription of the adenovirus genome.
Identification of proteins and protein domains that contact DNA within adenovirus nucleoprotein cores by ultraviolet light crosslinking of oligonucleotides <sup>32</sup>P-labelled in vivo
1986, Journal of Molecular Biology
A new approach to the identification of DNA binding proteins has been developed and used to study the DNA-protein interactions within the nucleoprotein core of subgroup C adenoviruses. Virions labelled in vivo with [³²P]orthophosphate were exposed to ultraviolet light and the DNA digested by chemical or enzymatic methods. Labelled phosphoamino acids of the virion phosphoproteins were selectively hydrolysed by alkali, permitting proteins crosslinked to DNA to be identified by virtue of their covalently attached, ³²P-labelled nucleotides. In parallel experiments, [³H]arginine-labelled virions were crosslinked by exposure to ultraviolet light and analysed by more conventional methods. The results indicate that proteins VII and V lie in close contact with viral DNA within the core. The compact arrangement of the nucleoprotein core appears to be capable of trapping protein VII molecules that are not covalently attached to DNA after exposure to ultraviolet light, suggesting that viral DNA might be wrapped around clusters of protein VII molecules. The domains of protein VII that lie in contact with DNA were identified by partial proteolytic mapping of the sites of covalent-attachment of the ³²P-labelled oligonucleotides. The implications of these data for the nature of the interactions that mediate the packaging of viral DNA within the nucleoprotein core of adenovirions are discussed.
Protein composition of adenovirus nucleoprotein complexes extracted from infected cells
1984, Virology
A viral nucleoprotein complex was extracted from the nuclei of human cells 20 hr after infection with adenovirus type 2 or several of its temperature-sensitive mutants. In its sedimentation property, density in CsCl, and digestion pattern with micrococcal nuclease, the complex resembled viral cores. The polypeptides V, PVII, IIK, and 36K were found associated with this complex which is formed prior to or in the absence of virus assembly. The results suggest that this nucleoprotein complex is a direct precursor to virus assembly.
Structure of adenovirus nucleoprotein core studied by circular dichroism and selective radiochemical labeling
1982, Biochemical and Biophysical Research Communications
The circular dichroic (CD) spectra of adenovirus DNA were compared within the intact virion and in deoxycholate and sarcosyl cores. It was found that the viral DNA was in highly condensed and packed conformation in both intact virus particles and deoxycholate cores, as in chromatin nucleosomes. Minor but significant CD spectral differences with eucaryotic chromatin indicated that the adenovirus nucleosome-like structures had specific conformational features. Adenovirus DNA only complexed with protein VII in sarcosyl cores elicited a CD spectrum reminiscent of a relaxed DNA, suggesting that the adenovirus sarcosyl cores were no longer organized in nucleosome subunits.
In parallel to CD analysis, the nature of the bonds involved in core structure wás investigated by in vitro selective labeling of some critical amino acids. ¹⁴C-dansyl was thus used to probe the accessibility of lysyl and tyrosyl residues in adenovirus core proteins. The data suggested that core protein V was associated with protein VII mainly via hydrophobic bonds, and had a major role in the maintainance of the nucleosomal structure of the viral core.
Comparison of the interactions of the adenovinis type 2 major core protein and its precursor with DNA
1986, Nucleic Acids Research

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The structure of adenovirion chromatin revealed by ultraviolet light-induced cross-linking

Abstract

Virology

Virol

Virology

J. Biol. Chem

J. Mol. Biol

J. Virol

J. Virol