PMID- 17372205
OWN - NLM
STAT- MEDLINE
DCOM- 20070514
LR  - 20181113
IS  - 0027-8424 (Print)
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Linking)
VI  - 104
IP  - 13
DP  - 2007 Mar 27
TI  - Insights on the role of nucleic acid/protein interactions in chaperoned nucleic 
      acid rearrangements of HIV-1 reverse transcription.
PG  - 5261-7
AB  - HIV-1 reverse transcription requires several nucleic acid rearrangement steps 
      that are "chaperoned" by the nucleocapsid protein (NC), including minus-strand 
      transfer, in which the DNA transactivation response element (TAR) is annealed to 
      the complementary TAR RNA region of the viral genome. These various rearrangement 
      processes occur in NC bound complexes of specific RNA and DNA structures. A major 
      barrier to the investigation of these processes in vitro has been the diversity 
      and heterogeneity of the observed nucleic acid/protein assemblies, ranging from 
      small complexes of only one or two nucleic acid molecules all the way up to 
      large-scale aggregates comprised of thousands of NC and nucleic acid molecules. 
      Herein, we use a flow chamber approach involving rapid NC/nucleic acid mixing to 
      substantially control aggregation for the NC chaperoned irreversible annealing 
      kinetics of a model TAR DNA hairpin sequence to the complementary TAR RNA 
      hairpin, i.e., to form an extended duplex. By combining the flow chamber approach 
      with a broad array of fluorescence single-molecule spectroscopy (SMS) tools 
      (FRET, molecule counting, and correlation spectroscopy), we have unraveled the 
      complex, heterogeneous kinetics that occur during the course of annealing. The 
      SMS results demonstrate that the TAR hairpin reactant is predominantly a single 
      hairpin coated by multiple NCs with a dynamic secondary structure, involving 
      equilibrium between a "Y" shaped conformation and a closed one. The data further 
      indicate that the nucleation of annealing occurs in an encounter complex that is 
      formed by two hairpins with one or both of the hairpins in the "Y" conformation.
FAU - Liu, Hsiao-Wei
AU  - Liu HW
AD  - Center for Nano and Molecular Science and Technology, University of Texas, 
      Austin, TX 78712, USA.
FAU - Zeng, Yining
AU  - Zeng Y
FAU - Landes, Christy F
AU  - Landes CF
FAU - Kim, Yoen Joo
AU  - Kim YJ
FAU - Zhu, Yongjin
AU  - Zhu Y
FAU - Ma, Xiaojing
AU  - Ma X
FAU - Vo, My-Nuong
AU  - Vo MN
FAU - Musier-Forsyth, Karin
AU  - Musier-Forsyth K
FAU - Barbara, Paul F
AU  - Barbara PF
LA  - eng
GR  - GM65056/GM/NIGMS NIH HHS/United States
GR  - F32 GM073534/GM/NIGMS NIH HHS/United States
GR  - GM65818/GM/NIGMS NIH HHS/United States
GR  - R01 GM065056/GM/NIGMS NIH HHS/United States
GR  - GM073534/GM/NIGMS NIH HHS/United States
GR  - R01 GM065818/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20070319
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Molecular Chaperones)
RN  - 0 (Nucleic Acids)
RN  - 0 (Oligonucleotides)
SB  - IM
MH  - Base Sequence
MH  - Fluorescence Resonance Energy Transfer
MH  - HIV-1/*metabolism
MH  - Kinetics
MH  - Molecular Chaperones
MH  - Molecular Sequence Data
MH  - Nucleic Acid Conformation
MH  - Nucleic Acids/*chemistry
MH  - Nucleocapsid/*chemistry
MH  - Oligonucleotides/chemistry
MH  - Protein Binding
MH  - Protein Conformation
MH  - Temperature
MH  - *Transcription, Genetic
MH  - *Transcriptional Activation
PMC - PMC1828707
COIS- The authors declare no conflict of interest.
EDAT- 2007/03/21 09:00
MHDA- 2007/05/15 09:00
PMCR- 2007/03/27
CRDT- 2007/03/21 09:00
PHST- 2007/03/21 09:00 [pubmed]
PHST- 2007/05/15 09:00 [medline]
PHST- 2007/03/21 09:00 [entrez]
PHST- 2007/03/27 00:00 [pmc-release]
AID - 0700166104 [pii]
AID - 5699 [pii]
AID - 10.1073/pnas.0700166104 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2007 Mar 27;104(13):5261-7. doi: 
      10.1073/pnas.0700166104. Epub 2007 Mar 19.