Kinetic mechanism for formation of the active, dimeric UvrD helicase-DNA complex.

The Journal of Biological Chemistry
Nasib K MalufTimothy M Lohman

Abstract

Escherichia coli UvrD protein is a 3' to 5' SF1 helicase required for DNA repair as well as DNA replication of certain plasmids. We have shown previously that UvrD can self-associate to form dimers and tetramers in the absence of DNA, but that a UvrD dimer is required to form an active helicase-DNA complex in vitro. Here we have used pre-steady state, chemical quenched flow methods to examine the kinetic mechanism for formation of the active, dimeric helicase-DNA complex. Experiments were designed to examine the steps leading to formation of the active complex, separate from the subsequent DNA unwinding steps. The results show that the active dimeric complex can form via two pathways. The first, faster path involves direct binding to the DNA substrate of a pre-assembled UvrD dimer (dimer path), whereas the second, slower path proceeds via sequential binding to the DNA substrate of two UvrD monomers (monomer path), which then assemble on the DNA to form the dimeric helicase. The rate-limiting step within the monomer pathway involves dimer assembly on the DNA. These results show that UvrD dimers that pre-assemble in the absence of DNA are intermediates along the pathway to formation of the functional dimeric UvrD helicase.

References

Apr 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·S H Northrup, H P Erickson
Oct 20, 1991·Journal of Molecular Biology·K L Chao, T M Lohman
Aug 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·G T RunyonT M Lohman
Dec 23, 1994·Science·A Sancar
Jan 1, 1994·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·S W MatsonJ W George
Jan 1, 1996·Annual Review of Biochemistry·P Modrich, R Lahue
Jan 1, 1996·Annual Review of Biochemistry·T M Lohman, K P Bjornson
Nov 28, 1996·Nature·H S SubramanyaD B Wigley
Apr 23, 1999·The Journal of Biological Chemistry·L E MechanicS W Matson
Nov 2, 1999·Journal of Molecular Biology·J A AliT M Lohman
Jan 13, 2000·Molecular Microbiology·C Bruand, S D Ehrlich
Sep 14, 2000·The Journal of Biological Chemistry·L E MechanicS W Matson
Jun 29, 2001·Journal of Molecular Biology·W ChengT M Lohman
Jan 16, 2003·Journal of Molecular Biology·Nasib K Maluf, Timothy M Lohman
Jan 16, 2003·Journal of Molecular Biology·Nasib K MalufTimothy M Lohman

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Citations

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May 23, 2013·Nature Communications·Kyung Suk LeeTaekjip Ha
Apr 17, 2008·Nature Reviews. Molecular Cell Biology·Timothy M LohmanColin G Wu
Mar 21, 2007·Molecular BioSystems·Alessandro Vindigni
Jun 6, 2012·Proceedings of the National Academy of Sciences of the United States of America·Kata SarlósMihály Kovács
Jan 24, 2012·The Journal of Biological Chemistry·Annamarie S CarterSteven W Matson
Aug 29, 2006·Nucleic Acids Research·Steven W Matson, Adam B Robertson
Dec 15, 2015·Nucleic Acids Research·Diana Constantinescu-AruxandeiMalcolm F White
Sep 30, 2010·The EMBO Journal·Eric J TomkoTimothy M Lohman
Nov 25, 2004·Journal of Molecular Biology·Christopher J FischerTimothy M Lohman
Oct 28, 2014·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Vitaliy Epshtein
May 30, 2015·FEMS Microbiology Letters·Hayat AlzahraniStephen J Forsythe
Apr 1, 2015·Nucleic Acids Research·Vessela PetrovaMichael M Cox
Jan 18, 2015·Current Opinion in Microbiology·Venu Kamarthapu, Evgeny Nudler
Nov 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Binh NguyenTimothy M Lohman
Nov 30, 2005·Journal of Physics. Condensed Matter : an Institute of Physics Journal·M D Betterton, F Jülicher
Jul 17, 2007·The Journal of Biological Chemistry·Anita Niedziela-MajkaTimothy M Lohman
May 7, 2014·The Journal of Biological Chemistry·Matthew J MeinersSteven W Matson
Sep 17, 2017·The Biochemical Journal·Kaustubh ShuklaGanesh Nagaraju
Aug 1, 2019·Proceedings of the National Academy of Sciences of the United States of America·Yerdos A OrdabayevTimothy M Lohman
Jan 28, 2021·International Journal of Molecular Sciences·Hiroaki Yokota
May 4, 2007·Biochemistry·Susan S DignamJohn David Dignam
Jun 4, 2021·Journal of Molecular Biology·Binh NguyenTimothy M Lohman
Aug 28, 2010·Biochemistry·Sergio Barranco-Medina, Roberto Galletto
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