HIV-1 Rev protein assembles on viral RNA one molecule at a time.
Oligomerization of the HIV-1 protein Rev on the Rev Response Element (RRE) regulates nuclear export of genomic viral RNA and partially spliced viral mRNAs encoding for structural proteins. Single-molecule fluorescence spectroscopy has been used to dissect the multistep assembly pathway of this essential ribonucleoprotein, revealing dynamic intermediates and the mechanism of assembly. Assembly is initiated by binding of Rev to a high-affinity site in stem-loop IIB of the RRE and proceeds rapidly by addition of single Rev monomers, facilitated by cooperative Rev-Rev interactions on the RRE. Dwell-time analysis of fluorescence trajectories recorded during individual Rev-RRE assembly reactions has revealed the microscopic rate constants for several of the Rev monomer binding and dissociation steps. The high-affinity binding of multiple Rev monomers to the RRE is achieved on a much faster timescale than reported in previous bulk kinetic studies of Rev-RRE association, indicating that oligomerization is an early step in complex assembly.
Pond
Stephanie J K
SJ
Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Ridgeway
William K
WK
Robertson
Rae
R
Wang
Jun
J
Millar
David P
DP
D000595:Amino Acid Sequence; D001483:Base Sequence; D015497:HIV-1 / Q000502:physiology*; D007700:Kinetics; D008856:Microscopy, Fluorescence; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D019065:Virus Assembly; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000502:physiology*
D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus; C514860:rev protein, Human Immunodeficiency Virus-1