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Journal of Virology, August 2001, p. 6865-6873, Vol. 75, No. 15
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.15.6865-6873.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Cytomegalovirus Basic Phosphoprotein (pUL32) Binds
to Capsids In Vitro through Its Amino One-Third
Michael K.
Baxter and
Wade
Gibson*
Virology Laboratories, Department of
Pharmacology and Molecular Sciences, The Johns Hopkins University
School of Medicine, Baltimore, Maryland 21205
Received 10 November 2000/Accepted 25 April 2001
The cytomegalovirus (CMV) basic phosphoprotein (BPP) is a component
of the tegument. It remains with the nucleocapsid fraction under
conditions that remove most other tegument proteins from the virion,
suggesting a direct and perhaps tight interaction with the capsid. As a
step toward localizing this protein within the molecular structure of
the virion and understanding its function during infection, we have
investigated the BPP-capsid interaction. In this report we present
evidence that the BPP interacts selectively, through its amino
one-third, with CMV capsids. Radiolabeled simian CMV (SCMV) BPP,
synthesized in vitro, bound to SCMV B-capsids, and C-capsids to a
lesser extent, following incubation with either isolated capsids or
lysates of infected cells. Human CMV (HCMV) BPP (pUL32) also bound to
SCMV capsids, and SCMV BPP likewise bound to HCMV capsids, indicating
that the sequence(s) involved is conserved between the two proteins.
Analysis of SCMV BPP truncation mutants localized the capsid-binding
region to the amino one-third of the molecule
the portion of BPP
showing the greatest sequence conservation between the SCMV and HCMV
homologs. This general approach may have utility in studying the
interactions of other proteins with conformation-dependent binding sites.
*
Corresponding author. Mailing address: Virology
Laboratories, Department of Pharmacology and Molecular Sciences, The
Johns Hopkins University School of Medicine, Baltimore, MD 21205. Phone: (410) 955-8680. Fax: (410) 955-3023. E-mail:
wgibson{at}jhmi.edu.
Journal of Virology, August 2001, p. 6865-6873, Vol. 75, No. 15
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.15.6865-6873.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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