HIV-1 resistance to the anti-HIV activity of a shRNA targeting a dual-coding region.
We generated a lymphoid cell line (Sup-T1-Rev/Env) that stably expresses a 19-bp short hairpin RNA (shRNA) targeting a conserved region of HIV-1 encoding for the Envelope and Rev proteins, which potently inhibited viral replication. However, continuous passage of HIV-1 in Sup-T1-Rev/Env generated virus mutants able to overcome the RNAi restriction. Sequence analysis of the emerging viruses showed that mutations were located at positions 5 and 17 of the target sequence. Both mutations are silent in the Env frame, but the mutation 5 generated an amino acid change (V47M) in the Rev reading frame. We have analyzed the impact of these two mutations on the RNAi mechanism, showing a more crucial role of the mutation 17 in the resistance to RNAi. We show that even targeting a conserved region of the HIV-1 genome involved in the biosynthesis of two essential genes, env and rev, the virus could evolve to escape by single point mutations in the target sequence, without a significant fitness cost.
Senserrich
Jordi
J
Retrovirology Laboratory irsiCaixa, Hospital Universitari Germans Trias i Pujol, Universitat Autònoma de Barcelona, 08916 Badalona, Spain.
Pauls
Eduardo
E
Armand-Ugón
Mercedes
M
Clotet-Codina
Imma
I
Moncunill
Gemma
G
Clotet
Bonaventura
B
Esté
Jose A
JA
D002460:Cell Line; D024882:Drug Resistance, Viral; D015752:Genes, env / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D009154:Mutation; D034622:RNA Interference; D014779:Virus Replication / Q000187:drug effects; D054299:env Gene Products, Human Immunodeficiency Virus / Q000235:genetics
D054299:env Gene Products, Human Immunodeficiency Virus