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PMID: 11932398 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Molecular organization of Mason-Pfizer monkey virus capsids assembled from Gag polyprotein in Escherichia coli.

Journal of virology ·Vol. 76 ·No. 9 ·2002-05-00 ·Pages 4321-30

Nermut MV, Bron P, Thomas D, Rumlova M, Ruml T, Hunter E

Abstract

We describe the results of a study by electron microscopy and image processing of Gag protein shells-immature capsids--of Mason-Pfizer monkey virus assembled in Escherichia coli from two truncated forms of the Gag precursor: Deltap4Gag, in which the C-terminal p4Gag was deleted, and Pro(-)CA.NC, in which the N-terminal peptides and proline 1 of the CA domain were deleted. Negative staining of capsids revealed small patches of holes forming a trigonal or hexagonal pattern most clearly visible on occasional tubular forms. The center-to-center spacing of holes in the network was 7.1 nm in Deltap4Gag capsids and 7.4 nm in Pro(-)CA.NC capsids. Image processing of Deltap4Gag tubes revealed a hexagonal network of holes formed by six subunits with a single subunit shared between rings. This organization suggests that the six subunits are contributed by three trimers of the truncated Gag precursor. Similar molecular organization was observed in negatively stained Pro(-)CA.NC capsids. Shadowed replicas of freeze-etched capsids produced by either construct confirmed the presence of a hexagonal network of holes with a similar center-to-center spacing. We conclude that the basic building block of the cage-like network is a trimer of the Deltap4Gag or Pro(-)CA.NC domains. In addition, our results point to a key role of structurally constrained CA domain in the trimeric interaction of the Gag polyprotein.

MeSH Terms
Animals Capsid/genetics,metabolism,ultrastructure Escherichia coli/genetics,metabolism Gene Products, gag/genetics,metabolism Image Processing, Computer-Assisted Mason-Pfizer monkey virus/genetics,metabolism Microscopy, Electron Virus Assembly
Chemicals
Gene Products, gag
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nermut Milan V
National Institute for Biological Standards and Control, South Mimms, Hertfordshire EN6 3QG, United Kingdom. [email protected]
Bron Patrick
Thomas Daniel
Rumlova Michaela
Ruml Tomas
Hunter Eric
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.76.9.4321-4330.2002
Email
[email protected]
Published
2002-05-00
Pages
4321-30
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC155098
Subset
IM
Grants
Wellcome Trust · United Kingdom
FIC NIH HHS · T37 TW000050 · United States
External Links
PubMed source DOI: 10.1128/jvi.76.9.4321-4330.2002 PMC Full Text View journal details
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