Resistance to RevM10 inhibition reflects a conformational switch in the HIV-1 Rev response element.
Nuclear export of certain HIV-1 mRNAs requires an interaction between the viral Rev protein and the Rev response element (RRE), a structured element located in the Env region of its RNA genome. This interaction is an attractive target for both drug design and gene therapy, exemplified by RevM10, a transdominant negative protein that, when introduced into host cells, disrupts viral mRNA export. However, two silent G->A mutations in the RRE (RRE61) confer RevM10 resistance, which prompted us to examine RRE structure using a novel chemical probing strategy. Variations in region III/IV/V of mutant RNAs suggest a stepwise rearrangement to RevM10 resistance. Mass spectrometry was used to directly assess Rev "loading" onto RRE and its variants, indicating that this is unaffected by RNA structural changes. Similarity in chemical footprints with mutant protein implicates additional host factors in RevM10 resistance.
Legiewicz
Michal
M
HIV Drug Resistance Program, National Cancer Institute, Frederick, MD 21702, USA.
Badorrek
Christopher S
CS
Turner
Kevin B
KB
Fabris
Daniele
D
Hamm
Tiffany E
TE
Rekosh
David
D
Hammarskjöld
Marie-Louise
ML
Le Grice
Stuart F J
SF
D001483:Base Sequence; D002460:Cell Line; D024882:Drug Resistance, Viral; D015752:Genes, env / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D006801:Humans; D013058:Mass Spectrometry; D008958:Models, Molecular; D009154:Mutation; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*
D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus
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