Characterization of human immunodeficiency virus type 1 dimeric RNA from wild-type and protease-defective virions.
We have characterized the dimeric genomic RNA in particles of both wild-type and protease (PR)-deficient human immunodeficiency virus type 1 (HIV-1). We found that the dimeric RNA isolated from PR- mutant virions has a lower mobility in nondenaturing gel electrophoresis than that from wild-type virions. It also dissociates into monomers at a lower temperature than the wild-type dimer. Thus, the dimer in PR- particles is in a conformation different from that in wild-type particles. These results are quite similar to recent findings on Moloney murine leukemia virus and suggest that a postassembly, PR-dependent maturation event is a common feature in genomic RNAs of retroviruses. We also measured the thermal stability of the wild-type and PR- dimeric RNAs under different ionic conditions. Both forms of the dimer were stabilized by increasing Na+ concentrations. However, the melting temperatures of the two forms were not significantly affected by the identity of the monovalent cation present in the incubation buffer. This observation is in contrast with recent reports on dimers formed in vitro from short segments of HIV-1 sequence: the latter dimers are specifically stabilized by K+ ions. K+ stabilization of dimers formed in vitro has been taken as evidence for the presence of guanine quartet structures. The results suggest that guanine quartets are not involved in the structure linking full-length, authentic genomic RNA of HIV-1 into a dimeric structure.
Fu
W
W
ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Maryland 21702-1201.
Gorelick
R J
RJ
Rein
A
A
D001704:Biopolymers; D016333:HIV Protease / Q000378:metabolism*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry*; D012965:Sodium Chloride; D013696:Temperature; D014771:Virion / Q000201:enzymology / Q000235:genetics
D001704:Biopolymers; D012367:RNA, Viral; D012965:Sodium Chloride; D016333:HIV Protease