Evidence for non-two-state kinetics in the nucleocapsid protein chaperoned opening of DNA hairpins.
In HIV-1 reverse transcription, the nucleocapsid protein, NC, induces secondary structure fluctuations in specific DNA and RNA hairpins. Time-resolved single-molecule fluorescence resonance energy transfer was used to study NC chaperoned opening of DNA hairpins over a broader range of conditions and in more depth than in previous studies. The experiments reveal a complex mechanism for secondary structure fluctuations with dynamic processes occurring over a wide time range, i.e., approximately 5 to >250 ms and with the involvement of long-lived intermediates. The dynamic role of DNA loop regions and NC binding/dissociation events are discussed.
Department of Chemistry and Biochemistry, Center for Nano and Molecular Science and Technology, University of Texas, Austin, Texas 78712, USA.
Gonzalo
G
Cosa
Yining
Y
Zeng
Hsiao-Wei
HW
Liu
Christy F
CF
Landes
Dmitrii E
DE
Makarov
Karin
K
Musier-Forsyth
Paul F
PF
Barbara
D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D018832:Molecular Chaperones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009690:Nucleic Acid Conformation; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D048348:Reverse Transcription
D018832:Molecular Chaperones; D019590:Nucleocapsid Proteins; D012313:RNA; D004247:DNA