Closing of the fingers domain generates motor forces in the HIV reverse transcriptase

Hailong Lu; Jed Macosko; Diana Habel-Rodriguez; Rebecca W Keller; James A Brozik; David J Keller

doi:10.1074/jbc.M407193200

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Closing of the fingers domain generates motor forces in the HIV reverse transcriptase

Journal article

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Peer reviewed

Closing of the fingers domain generates motor forces in the HIV reverse transcriptase

Hailong Lu, Jed Macosko, Diana Habel-Rodriguez, Rebecca W Keller, James A Brozik and David J Keller

The Journal of biological chemistry, Vol.279(52), pp.54529-54532

12/24/2004

DOI: https://doi.org/10.1074/jbc.M407193200

Handle:

https://hdl.handle.net/2376/106170

PMID: 15385563

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Abstract

Biotinylation

Models, Molecular

Templates, Genetic

Structure-Activity Relationship

DNA - metabolism

Chemistry, Physical

HIV Reverse Transcriptase - metabolism

DNA - chemistry

Chemical Phenomena

Protein Conformation

Catalysis

Kinetics

Binding Sites

HIV Reverse Transcriptase - chemistry

Polymers - metabolism

Using the force sensor of an atomic force microscope, motor forces of the human immunodeficiency virus-1 reverse transcriptase were measured during active replication of a short DNA transcript. At low load forces the polymerase is mechanically slowed, whereas at high force (approximately 15 piconewton) it stalls. From recordings of estimated polymerase turnover velocity versus load force, an approximate force-velocity curve has been constructed. The shape of the curve suggests that load force strongly inhibits the rate-limiting step of the polymerase turnover cycle and that the combined effect of load on all steps involves an effective motion of about 1.6 nm. Earlier results from pre-steady-state kinetics experiments have identified the rate-limiting step as the closing of the fingers domain to form a tight catalytic complex. Together these findings indicate that the closing of the fingers domain is a major force-generating step for human immunodeficiency virus reverse transcriptase and, by extension, for all DNA polymerase machines.

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Details

Title: Closing of the fingers domain generates motor forces in the HIV reverse transcriptase
Creators: Hailong Lu - Department of Chemistry, The University of New Mexico, Albuquerque, New Mexico 87106, USA
Jed Macosko
Diana Habel-Rodriguez
Rebecca W Keller
James A Brozik
David J Keller
Publication Details: The Journal of biological chemistry, Vol.279(52), pp.54529-54532
Academic Unit: Chemistry, Department of
Publisher: United States
Grant note: GM63808 / NIGMS NIH HHS
Identifiers: 99900546905101842
Language: English
Resource Type: Journal article