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N‐myristoyltransferase‐2: a potential regulator of Nef‐mediated HIV pathogenesis
Author(s) -
Seaton Kelly Elizabeth,
French Kevin,
Smith Charles
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a443
Subject(s) - myristoylation , microbiology and biotechnology , regulator , biology , infectivity , recombinant dna , viral replication , human immunodeficiency virus (hiv) , transfection , virology , virus , cell culture , biochemistry , phosphorylation , genetics , gene
N‐terminal myristoylation of the HIV proteins Gag and Nef is critical to retroviral replication, virulence and budding. Mammals express two N‐myristoyltransferases (NMT1 and NMT2), but it is not yet known if they have separate or overlapping activities toward the HIV proteins. Elucidation of the roles of the NMTs in Gag and Nef myristoylation would provide mechanistic insight into the biology of HIV and identify potential targets for new anti‐AIDS drugs. Kinetics for Gag and Nef myristoylation were determined using peptides corresponding to their N‐termini and recombinant human NMT1 or NMT2. Results indicate that both isozymes have lower K m s for Nef than Gag; while NMT2 has a 6‐fold higher catalytic efficiency toward Nef than does NMT1. Confocal microscopy indicated that selective depletion of NMT1 had minimal affect on the intracellular distribution of Nef‐sgGFP. In contrast, ablation of NMT2 altered the distribution of Nef‐sgGFP to a pattern similar to a G2→A Nef‐sgGFP construct. We are currently examining the effects of selective NMT depletion (using siRNA or pharmacologic agents) on Nef function in intact HIV‐infected cells. Together, these findings indicate that Nef is preferentially myristoylated by NMT2 such that selective inhibition of NMT2 may provide a novel means of blocking HIV infectivity. [2R01CA75248]