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PMID: 7966591 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The p2 domain of human immunodeficiency virus type 1 Gag regulates sequential proteolytic processing and is required to produce fully infectious virions.

Journal of virology ·Vol. 68 ·No. 12 ·1994-12-00 ·Pages 8017-27

Pettit SC, Moody MD, Wehbie RS, Kaplan AH, Nantermet PV, Klein CA, Swanstrom R

Abstract

The proteolytic processing sites of the human immunodeficiency virus type 1 (HIV-1) Gag precursor are cleaved in a sequential manner by the viral protease. We investigated the factors that regulate sequential processing. When full-length Gag protein was digested with recombinant HIV-1 protease in vitro, four of the five major processing sites in Gag were cleaved at rates that differ by as much as 400-fold. Three of these four processing sites were cleaved independently of the others. The CA/p2 site, however, was cleaved approximately 20-fold faster when the adjacent downstream p2/NC site was blocked from cleavage or when the p2 domain of Gag was deleted. These results suggest that the presence of a C-terminal p2 tail on processing intermediates slows cleavage at the upstream CA/p2 site. We also found that lower pH selectively accelerated cleavage of the CA/p2 processing site in the full-length precursor and as a peptide primarily by a sequence-based mechanism rather than by a change in protein conformation. Deletion of the p2 domain of Gag results in released virions that are less infectious despite the presence of the processed final products of Gag. These findings suggest that the p2 domain of HIV-1 Gag regulates the rate of cleavage at the CA/p2 processing site during sequential processing in vitro and in infected cells and that p2 may function in the proper assembly of virions.

Related Genes
gag
MeSH Terms
Amino Acid Sequence Base Sequence Blotting, Western Cloning, Molecular Gene Products, gag/biosynthesis,metabolism Genes, gag HIV-1/genetics,metabolism,pathogenicity HeLa Cells Humans Kinetics Molecular Sequence Data Mutagenesis, Site-Directed Oligodeoxyribonucleotides Protein Biosynthesis Protein Precursors/metabolism Protein Processing, Post-Translational RNA, Messenger/biosynthesis,metabolism Recombinant Proteins/biosynthesis,metabolism Transfection Virion/genetics,metabolism,pathogenicity
Chemicals
Gene Products, gag Oligodeoxyribonucleotides Protein Precursors RNA, Messenger Recombinant Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Pettit S C
UNC Lineberger Comprehensive Cancer Center, Chapel Hill.
Moody M D
Wehbie R S
Kaplan A H
Nantermet P V
Klein C A
Swanstrom R
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.68.12.8017-8027.1994
Published
1994-12-00
Pages
8017-27
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237265
Subset
IM
Grants
NIAID NIH HHS · F32-AI08400 · United States
NIAID NIH HHS · F32-AI08767 · United States
NIAID NIH HHS · R01-AI25321 · United States
External Links
PubMed source DOI: 10.1128/JVI.68.12.8017-8027.1994 PMC Full Text View journal details
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