PMID: 8955285Dec 1, 1996Paper

Interaction of Escherichia coli primase with a phage G4ori(c)-E. coli SSB complex

Journal of Bacteriology
W Sun, G N Godson

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

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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 5, 2006·Journal of Bacteriology·Anna Rodina, G Nigel Godson
Jun 15, 1999·Journal of Bacteriology·W SunG N Godson
Mar 13, 2001·Journal of Molecular Biology·D SikderV Nagaraja
Aug 15, 2021·Journal of Molecular Biology·Christina N GirammaJue D Wang

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