Binding of trans-dominant mutant Rev protein of human immunodeficiency virus type 1 to the cis-acting Rev-responsive element does not affect the fate of viral mRNA.
The binding of Rev protein of human immunodeficiency virus type 1 (HIV-1) to the cis-acting Rev-responsive element (RRE) was compared to the binding of a trans-dominant Rev mutant. RevBL, which inhibits Rev function. Rev and RevBL expressed in bacteria were purified and shown to bind in vitro to the RRE with similar affinities. The study of the RRE mutants indicated that Rev and RevBL bind to the same target within the RRE in vitro and in vivo. In vivo experiments demonstrated that RevBL did not increase the steady-state levels of HIV-1 mRNA or protein. These experiments suggested that additional cellular factors interacting with Rev but not with RevBL are necessary for function. The Rex protein of human T-cell leukemia virus type I (HTLV-I) is similar to Rev and acts through a sequence named Rex-responsive element (RXRE) located in the long terminal repeat of HTLV-I. We examined the function of RevBL on a hybrid mRNA molecule containing both the RRE and RXRE. While RevBL prevented Rev function, it did not affect Rex function on the mRNA containing either the RXRE or both the RRE and RXRE. Therefore, binding of RevBL to the RRE had neither positive nor negative effects on the mRNA, since this mRNA could be efficiently utilized in the presence of a functional Rex-RXRE interaction. The results obtained in vivo and in vitro strongly suggest that RevBL inhibits Rev function by binding to the same site as Rev and preventing Rev binding and function.
Benko
D M
DM
ABL-Basic Research Program, National Cancer Institute-FCRDC, MD 21702-1201.
Robinson
R
R
Solomin
L
L
Mellini
M
M
Felber
B K
BK
Pavlakis
G N
GN
D015967:Gene Expression Regulation, Viral; D015683:Gene Products, gag / Q000096:biosynthesis / Q000235:genetics; D015694:Gene Products, rev / Q000235:genetics*; D015757:Genes, rev; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D009154:Mutation; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000378:metabolism*; D012367:RNA, Viral; D012045:Regulatory Sequences, Nucleic Acid; D054321:rev Gene Products, Human Immunodeficiency Virus
D015683:Gene Products, gag; D015694:Gene Products, rev; D012333:RNA, Messenger; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus