Functional and structural heterogeneity of the DNA binding site of the Escherichia coli primary replicative helicase DnaB protein.

The Journal of Biological Chemistry
M J JezewskaW Bujalowski

Abstract

The structure-function relationship within the DNA binding site of the Escherichia coli replicative helicase DnaB protein was studied using nuclease digestion, quantitative fluorescence titration, centrifugation, and fluorescence energy transfer techniques. Nuclease digestion of the enzyme-single-stranded DNA (ssDNA) complexes reveals large structural heterogeneity within the binding site. The total site is built of two subsites differing in structure and affinity, although both occlude approximately 10 nucleotides. ssDNA affinity for the strong subsite is approximately 3 orders of magnitude higher than that for the weak subsite. Fluorescence energy transfer experiments provide direct proof that the DnaB hexamer binds ssDNA in a single orientation, with respect to the polarity of the sugar-phosphate backbone. This is the first evidence of directional binding to ssDNA of a hexameric helicase in solution. The strong binding subsite is close to the small 12-kDa domains of the DnaB hexamer and occludes the 5'-end of the ssDNA. The strict orientation of the helicase on ssDNA indicates that, when the enzyme approaches the replication fork, it faces double-stranded DNA with its weak subsite. The data indicate that the different bindin...Continue Reading

References

Nov 4, 1976·Journal of Molecular Biology·M T RecordP De Haseth
Mar 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·S WicknerJ Hurwitz
May 1, 1995·Journal of Structural Biology·M C San MartinJ M Carazo
Apr 25, 1995·Proceedings of the National Academy of Sciences of the United States of America·E H EgelmanS S Patel
May 31, 1996·Journal of Molecular Biology·X YuE H Egelman
Jan 1, 1996·Annual Review of Biochemistry·T M Lohman, K P Bjornson
Sep 1, 1971·Proceedings of the National Academy of Sciences of the United States of America·C R Cantor, P Pechukas

❮ Previous
Next ❯

Citations

Mar 14, 2003·Trends in Biochemical Sciences·Wlodzimierz Bujalowski
May 30, 2002·Nucleic Acids Research·Silvia AyoraJuan C Alonso
Dec 6, 2002·Nucleic Acids Research·María I Martínez-JiménezJuan C Alonso
Aug 9, 2006·Nucleic Acids Research·Sandrine CastellaCyril M Sanders
Sep 9, 2005·Microbiology and Molecular Biology Reviews : MMBR·Cameron NeylonNicholas E Dixon
Feb 20, 2013·Biochemistry·Michal R SzymanskiWlodzimierz Bujalowski
Feb 20, 2013·Biochemistry·Michal R SzymanskiWlodzimierz Bujalowski
Oct 14, 2014·Journal of Molecular Biology·Jan-Gert BrüningPeter McGlynn
Mar 11, 2008·Current Opinion in Structural Biology·Eric J Enemark, Leemor Joshua-Tor
Oct 2, 2012·Cell·Ornchuma ItsathitphaisarnThomas A Steitz
Feb 5, 2010·Molecular Cell·Magdalena Makowska-Grzyska, Jon M Kaguni
Aug 31, 2000·Annual Review of Biochemistry·S S Patel, K M Picha
Aug 27, 2016·Frontiers in Molecular Biosciences·Katarzyna E WegrzynIgor Konieczny
Jul 23, 2005·IUBMB Life·Patrick M SchaefferNicholas E Dixon
Nov 2, 1999·Journal of Molecular Biology·H GeorgeS C West
Jan 31, 2018·Nature Structural & Molecular Biology·Michael E O'Donnell, Huilin Li
Sep 25, 2017·The Journal of Biological Chemistry·Sean M CarneyMichael A Trakselis
Sep 24, 2002·The Journal of Biological Chemistry·Alexander I AlexandrovNicholas R Cozzarelli
Mar 26, 2002·The Journal of Biological Chemistry·Maria J JezewskaWlodzimierz Bujalowski
Oct 14, 2000·Journal of Molecular Biology·L E DonateJ M Carazo
Dec 5, 1998·Journal of Molecular Biology·M J JezewskaW Bujalowski

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.