HIV-1-associated PKA acts as a cofactor for genome reverse transcription.
Retrovirology
; 10: 157, 2013 Dec 17.
Article
em En
| MEDLINE
| ID: mdl-24344931
ABSTRACT
BACKGROUND:
Host cell proteins, including cellular kinases, are embarked into intact HIV-1 particles. We have previously shown that the Cα catalytic subunit of cAMP-dependent protein kinase is packaged within HIV-1 virions as an enzymatically active form able to phosphorylate a synthetic substrate in vitro (Cartier et al. J. Biol. Chem. 27835211 (2003)). The present study was conceived to investigate the contribution of HIV-1-associated PKA to the retroviral life cycle.RESULTS:
NL4.3 viruses were produced from cells cultured in the presence of PKA inhibitors H89 (H89-NL4.3) or Myr-PKI (PKI-NL4.3) and analyzed for viral replication. Despite being mature and normally assembled, and containing expected levels of genomic RNA and RT enzymatic activity, such viruses showed poor infectivity. Indeed, infection generated reduced amounts of strong-strop minus strand DNA, while incoming RNA levels in target cells were unaffected. Decreased cDNA synthesis was also evidenced in intact H89-NL4.3 and PKI-NL4.3 cell free particles using endogenous reverse transcription (ERT) experiments. Moreover, similar defects were reproduced when wild type NL4.3 particles preincubated with PKA inhibitors were subjected to ERT reactions.CONCLUSIONS:
Altogether, our results indicate that HIV-1-associated PKA is required for early reverse transcription of the retroviral genome both in cell free intact viruses and in target cells. Accordingly, virus-associated PKA behaves as a cofactor of an intraviral process required for optimal reverse transcription and for early post-entry events.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
HIV-1
/
Integração Viral
/
Proteínas Quinases Dependentes de AMP Cíclico
/
Transcrição Reversa
/
Interações Hospedeiro-Patógeno
Tipo de estudo:
Risk_factors_studies
Limite:
Humans
Idioma:
En
Revista:
Retrovirology
Assunto da revista:
VIROLOGIA
Ano de publicação:
2013
Tipo de documento:
Article