Abstract
We earlier reported that Escherichia coli single-stranded DNA-binding protein (SSB) bound in a fixed position to the stem-loop structure of the origin of complementary DNA strand synthesis in phage G4 (G4ori(c)), leaving stem-loop I and the adjacent 5' CTG 3', the primer RNA initiation site, as an SSB-free region (W. Sun and G. N. Godson, J. Biol. Chem. 268:8026-8039, 1993). Using a small 278-nucleotide (nt) G4ori(c) single-stranded DNA fragment that supported primer RNA synthesis, we now demonstrate by gel shift that E. coli primase can stably interact with the SSB-G4ori(c) complex. This stable interaction requires Mg2+ for specificity. At 8 mM Mg2+, primase binds to an SSB-coated 278-nt G4ori(c) fragment but not to an SSB-coated control 285-nt LacZ ss-DNA fragment. In the absence of Mg2+, primase binds to both SSB-coated fragments and gives a gel shift. T4 gene 32 protein cannot substitute for E. coli SSB in this reaction. Stable interaction of primase with naked G4ori(c). single-stranded DNA was not observed. DNase I and micrococcal nuclease footprinting, of both 5' and 3' 32P-labeled DNA, demonstrated that primase interacts with two regions of G4ori(c): one covering stem-loop I and the 3' sequence flanking stem-loop I which...Continue Reading
References
Jul 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·S Wickner
Mar 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·J C FiddesG N Godson
Jul 1, 1978·Proceedings of the National Academy of Sciences of the United States of America·J Sims, D Dressler
Nov 16, 1978·Nature·G N GodsonJ C Fiddes
Sep 11, 1992·Biochemistry·W C SuhM T Record
Aug 25, 1990·Nucleic Acids Research·H HiasaG N Godson
May 1, 1991·Molecular & General Genetics : MGG·K Yoda, T Okazaki
Apr 1, 1991·Gene·G N Godson
Jun 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·A van der EndeA Kornberg
Oct 3, 1985·Biochimica Et Biophysica Acta·H Sakai, G N Godson
Jun 15, 1987·European Journal of Biochemistry·Y F Roth
Jan 1, 1987·Journal of Cell Science. Supplement·A Kornberg
Oct 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·M S WoldR McMacken
Feb 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·J Sims, E W Benz
Jun 30, 1995·The Journal of Biological Chemistry·A A Mustaev, G N Godson
Dec 26, 1995·Biochemistry·T M Urlacher, M A Griep
Jul 5, 1996·Science·E ZaychikovA Mustaev
Citations
Jun 8, 2001·Annual Review of Biochemistry·D N Frick, C C Richardson
Aug 6, 2002·The Journal of Biological Chemistry·Kevin M Carr, Jon M Kaguni
Mar 21, 2009·Annual Review of Biochemistry·Samir M Hamdan, Charles C Richardson
Feb 23, 2018·Antibiotics·Zorik ChilingaryanAaron J Oakley
Jun 15, 2005·Annual Review of Biochemistry·Aaron Johnson, Mike O'Donnell
Aug 29, 2006·Nucleic Acids Research·Jacob E Corn, James M Berger
May 3, 2008·Molecular Microbiology·Scott A KoepsellMark A Griep
May 5, 2006·Journal of Bacteriology·Anna Rodina, G Nigel Godson
May 20, 2020·Molecular Microbiology·Christine WolakJames L Keck
Jun 15, 1999·Journal of Bacteriology·W SunG N Godson
May 9, 1998·Journal of Molecular Biology·W Sun, G N Godson
Mar 13, 2001·Journal of Molecular Biology·D SikderV Nagaraja
Aug 15, 2021·Journal of Molecular Biology·Christina N GirammaJue D Wang
Dec 20, 2008·Journal of Molecular Biology·Ruchika Sharma, Desirazu N Rao
Jul 17, 2012·Structure·Richard U RymerJames M Berger