A poxvirus host range protein, CP77, binds to a cellular protein, HMG20A, and regulates its dissociation from the vaccinia virus genome in CHO-K1 cells

Jye Chian Hsiao, Chien Chiang Chao, Ming Jer Young, Yu Tai Chang, Er Chieh Cho, Wen Chang

Research output: Contribution to journalArticle

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Abstract

Vaccinia virus does not grow in Chinese hamster ovary (CHO-K1) cells in the absence of a viral host range factor, cowpox protein CP77. In this study, CP77 was fused to the C terminus of green fluorescence protein (GFP-CP77) and a series of nested deletion mutants of GFP-CP77 was constructed for insertion into a vaccinia virus host range mutant, VV-hr, and expressed from a viral early promoter. Deletion mapping analyses demonstrated that the N-terminal 352 amino acids of CP77 were sufficient to support vaccinia virus growth in CHO-K1 cells, whereas the C-terminal residues 353 to 668 were dispensable. In yeast two-hybrid analyses, CP77 bound to a cellular protein, HMG20A, and GST pulldown analyses showed that residues 1 to 234 of CP77 were sufficient for this interaction. After VV-hr virus infection of CHO-K1 cells, HMG20A was translocated from the nucleus to viral factories and bound to the viral genome via the HMG box region. In control VV-hr-infected CHO-K1 cells, binding of HMG20A to the viral genome persisted from 2 to 8 h postinfection (h p.i.); in contrast, when CP77 was expressed, the association of HMG20A with viral genome was transient, with little HMG20A remaining bound at 8 h p.i. This indicates that dissociation of HMG20A from viral factories correlates well with CP77 host range activity in CHO-K1 cells. Finally, in cells expressing a CP77 deletion protein (amino acids 277 to 668) or a ΔANK5 mutant that did not support vaccinia virus growth and did not contain the HMG20A binding site, HMG20A remained bound to viral DNA, demonstrating that the binding of CP77 to HMG20A is essential for its host range function. In summary, our data revealed that a novel cellular protein, HMG20A, the dissociation of which from viral DNA is regulated by CP77, providing the first cellular target regulated by viral host range CP77 protein.

Original languageEnglish
Pages (from-to)7714-7728
Number of pages15
JournalJournal of Virology
Volume80
Issue number15
DOIs
Publication statusPublished - Aug 2006

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Poxviridae
Vaccinia virus
CHO Cells
Host Specificity
host range
Genome
genome
Viral Genome
Proteins
proteins
Viral DNA
cells
factories
mutants
Cowpox
Amino Acids
amino acids
Chinese hamsters
DNA
Virus Diseases

ASJC Scopus subject areas

  • Immunology

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A poxvirus host range protein, CP77, binds to a cellular protein, HMG20A, and regulates its dissociation from the vaccinia virus genome in CHO-K1 cells. / Hsiao, Jye Chian; Chao, Chien Chiang; Young, Ming Jer; Chang, Yu Tai; Cho, Er Chieh; Chang, Wen.

In: Journal of Virology, Vol. 80, No. 15, 08.2006, p. 7714-7728.

Research output: Contribution to journalArticle

Hsiao, Jye Chian ; Chao, Chien Chiang ; Young, Ming Jer ; Chang, Yu Tai ; Cho, Er Chieh ; Chang, Wen. / A poxvirus host range protein, CP77, binds to a cellular protein, HMG20A, and regulates its dissociation from the vaccinia virus genome in CHO-K1 cells. In: Journal of Virology. 2006 ; Vol. 80, No. 15. pp. 7714-7728.
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