Identification of a DNA primase template tracking site redefines the geometry of primer synthesis

Nature Structural & Molecular Biology
Jacob E CornJames M Berger

Abstract

Primases are essential RNA polymerases required for the initiation of DNA replication, lagging strand synthesis and replication restart. Many aspects of primase function remain unclear, including how the enzyme associates with a moving nucleic acid strand emanating from a helicase and orients primers for handoff to replisomal components. Using a new screening method to trap transient macromolecular interactions, we determined the structure of the Escherichia coli DnaG primase catalytic domain bound to single-stranded DNA. The structure reveals an unanticipated binding site that engages nucleic acid in two distinct configurations, indicating that it serves as a nonspecific capture and tracking locus for template DNA. Bioinformatic and biochemical analyses show that this evolutionarily constrained region enforces template polarity near the active site and is required for primase function. Together, our findings reverse previous proposals for primer-template orientation and reconcile disparate studies to re-evaluate replication fork organization.

References

Mar 31, 1998·Current Opinion in Structural Biology·C A Brautigam, T A Steitz
Jun 15, 1999·Journal of Bacteriology·W SunG N Godson
Dec 22, 1999·Science·G M Cheetham, T A Steitz
Mar 31, 2000·Science·J L KeckJ M Berger
Jun 8, 2001·Annual Review of Biochemistry·D N Frick, C C Richardson
Aug 23, 2001·Proceedings of the National Academy of Sciences of the United States of America·N A BakerJ A McCammon
Sep 14, 2002·Proceedings of the National Academy of Sciences of the United States of America·Seung-Joo Lee, Charles C Richardson
Oct 7, 2003·Annual Review of Biochemistry·Gregory L Verdine, Derek P G Norman
Nov 26, 2003·Molecular Cell·Eric A TothTom Ellenberger
Mar 3, 2004·Acta Crystallographica. Section D, Biological Crystallography·Laurent C StoroniRandy J Read
Mar 16, 2004·Cell·Andrew I ShulmanRama Ranganathan
Mar 23, 2004·Nucleic Acids Research·Robert C Edgar
May 1, 1997·Acta Crystallographica. Section D, Biological Crystallography·G N MurshudovE J Dodson
Oct 22, 2004·Journal of Synchrotron Radiation·Alastair A MacDowellHoward A Padmore
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Nov 16, 2005·Molecular Cell·Jacob E CornJames M Berger
Feb 3, 2006·Nature·Ryan C Heller, Kenneth J Marians
Apr 28, 2006·The EMBO Journal·Udi QimronCharles C Richardson
May 5, 2006·Journal of Bacteriology·Anna Rodina, G Nigel Godson
Apr 25, 2007·Nucleic Acids Research·Ian W DavisDavid C Richardson

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Citations

Feb 6, 2008·Nature Structural & Molecular Biology·Kenneth J Marians
May 7, 2009·Proceedings of the National Academy of Sciences of the United States of America·Sebastian GeibelWolfram Saenger
Jul 21, 2010·Proceedings of the National Academy of Sciences of the United States of America·Sivaraja VaithiyalingamWalter J Chazin
May 5, 2010·Proceedings of the National Academy of Sciences of the United States of America·Bin ZhuCharles C Richardson
Sep 17, 2010·The Journal of Biological Chemistry·Brian R BowmanGregory L Verdine
Apr 19, 2012·The Journal of Biological Chemistry·Charisse M CrenshawGregory L Verdine
Jan 11, 2012·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Dong-Won ImYong-Seok Heo
Apr 16, 2013·Journal of Bacteriology·Syed Arif Abdul RehmanS Gourinath
May 29, 2010·Microbiology and Molecular Biology Reviews : MMBR·Andrew RobinsonNicholas E Dixon
Dec 1, 2015·Critical Reviews in Biochemistry and Molecular Biology·Laurie S Kaguni, Marcos T Oliveira
Jan 30, 2013·Journal of Molecular Biology·Robert J BauerMichael A Trakselis
Aug 23, 2011·Current Opinion in Chemical Biology·Jon M Kaguni
Aug 26, 2011·Current Opinion in Chemical Biology·Smita S PatelDivya Nandakumar
May 12, 2011·Mutation Research·Marc BicharaIain B Lambert
Oct 3, 2013·DNA Repair·Shawn C Massoni, Steven J Sandler
Nov 19, 2013·Journal of Molecular Biology·Sivaraja VaithiyalingamWalter J Chazin
Apr 1, 2015·Nucleic Acids Research·Pablo Fernández-MillánMaria Solà
Feb 5, 2010·Molecular Cell·Magdalena Makowska-Grzyska, Jon M Kaguni
Feb 4, 2010·Journal of Molecular Biology·Zhongfeng ZuoMichael A Trakselis
Jan 12, 2017·Genes·Katie H Jameson, Anthony J Wilkinson
Oct 27, 2017·The Journal of Biological Chemistry·Magdalena M FelczakJon M Kaguni
Dec 1, 2018·Protein Science : a Publication of the Protein Society·Sergio E MartinezEddy Arnold
Apr 5, 2019·Protein Science : a Publication of the Protein Society·Aaron J Oakley
May 26, 2012·Molecular BioSystems·Roger J HeinzePeter Friedhoff
Apr 8, 2017·Scientific Reports·Yingqin ZhouGanggang Wang
Oct 5, 2019·Current Opinion in Structural Biology·Jan BergschGeorg Lipps
Jun 23, 2009·Biochimica Et Biophysica Acta·Robert D Kuchta, Gudrun Stengel
Sep 11, 2012·Structure·Robyn E Mansfield, Nicholas E Dixon

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