Characterization of a peptide domain within the GB virus C NS5A phosphoprotein that inhibits HIV replication.
BACKGROUND
GBV-C infection is associated with prolonged survival in HIV-infected people and GBV-C inhibits HIV replication in co-infection models. Expression of the GBV-C nonstructural phosphoprotein 5A (NS5A) decreases surface levels of the HIV co-receptor CXCR4, induces the release of SDF-1 and inhibits HIV replication in Jurkat CD4+ T cell lines.


RESULTS
Jurkat cell lines stably expressing NS5A protein and peptides were generated and HIV replication in these cell lines assessed. HIV replication was significantly inhibited in all cell lines expressing NS5A amino acids 152-165. Substitution of an either alanine or glycine for the serine at position 158 (S158A or S158G) resulted in a significant decrease in the HIV inhibitory effect. In contrast, substituting a phosphomimetic amino acid (glutamic acid; S158E) inhibited HIV as well as the parent peptide. HIV inhibition was associated with lower levels of surface expression of the HIV co-receptor CXCR4 and increased release of the CXCR4 ligand, SDF-1 compared to control cells. Incubation of CD4+ T cell lines with synthetic peptides containing amino acids 152-167 or the S158E mutant peptide prior to HIV infection resulted in HIV replication inhibition compared to control peptides.


CONCLUSIONS
Expression of GBV-C NS5A amino acids 152-165 are sufficient to inhibit HIV replication in vitro, and the serine at position 158 appears important for this effect through either phosphorylation or structural changes in this peptide. The addition of synthetic peptides containing 152-167 or the S158E substitution to Jurkat cells resulted in HIV replication inhibition in vitro. These data suggest that GBV-C peptides or a peptide mimetic may offer a novel, cellular-based approach to antiretroviral therapy.
Xiang
Jinhua
J
Iowa City VA Medical Center and the University of Iowa, Iowa City, Iowa, United States of America.
McLinden
James H
JH
Chang
Qing
Q
Jordan
Emma L
EL
Stapleton
Jack T
JT
D019380:Anti-HIV Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D015536:Down-Regulation; D029407:GB virus C; D006678:HIV / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D010455:Peptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010766:Phosphorylation; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D014779:Virus Replication / Q000187:drug effects*
D019380:Anti-HIV Agents; D010455:Peptides; D010750:Phosphoproteins; D017361:Viral Nonstructural Proteins; C532378:nonstructural phosphoprotein 5A, GB virus type C