Escherichia coli DNA polymerase II is stimulated by DNA polymerase III holoenzyme auxiliary subunits.
DNA polymerase III of Escherichia coli requires multiple auxiliary factors to enable it to serve as a replicative complex. We demonstrate that auxiliary components of the DNA polymerase III holoenzyme, the gamma delta complex and beta subunit, markedly stimulate DNA polymerase II on long single-stranded templates. DNA polymerase II activity is enhanced by single-stranded DNA binding protein, but the stimulation by gamma delta and beta can be observed either in the absence or presence of single-stranded DNA binding protein. In contrast with DNA polymerase III, the requirement of DNA polymerase II for gamma delta cannot be bypassed by large excesses of the beta subunit at low ionic strength in the absence of the single-stranded DNA binding protein. The product of the DNA polymerase II-gamma delta-beta reaction on a uniquely primed single-stranded circle is of full template length; the reconstituted enzyme apparently is incapable of strand displacement synthesis. The possible biological implications of these observations are discussed.
Hughes
A J
AJ
Department of Biochemistry, Biophysics, and Genetics, University of Colorado Health Sciences Center, Denver 80262.
Bryan
S K
SK
Chen
H
H
Moses
R E
RE
McHenry
C S
CS
D004257:DNA Polymerase II / Q000378:metabolism*; D004258:DNA Polymerase III / Q000378:metabolism*; D004261:DNA Replication; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D009994:Osmolar Concentration; D013329:Structure-Activity Relationship; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D004257:DNA Polymerase II; D004258:DNA Polymerase III