The DNA polymerase III holoenzyme contains γ and is not a trimeric polymerase

Nucleic Acids Research
Paul R DohrmannCharles S McHenry

Abstract

There is widespread agreement that the clamp loader of the Escherichia coli replicase has the composition DnaX3δδ'χψ. Two DnaX proteins exist in E. coli, full length τ and a truncated γ that is created by ribosomal frameshifting. τ binds DNA polymerase III tightly; γ does not. There is a controversy as to whether or not DNA polymerase III holoenzyme (Pol III HE) contains γ. A three-τ form of Pol III HE would contain three Pol IIIs. Proponents of the three-τ hypothesis have claimed that γ found in Pol III HE might be a proteolysis product of τ. To resolve this controversy, we constructed a strain that expressed only τ from a mutated chromosomal dnaX. γ containing a C-terminal biotinylation tag (γ-C(tag)) was provided in trans at physiological levels from a plasmid. A 2000-fold purification of Pol III* (all Pol III HE subunits except β) from this strain contained one molecule of γ-C(tag) per Pol III* assembly, indicating that the dominant form of Pol III* in cells is Pol III2τ2 γδδ'χψ. Revealing a role for γ in cells, mutants that express only τ display sensitivity to ultraviolet light and reduction in DNA Pol IV-dependent mutagenesis associated with double-strand-break repair, and impaired maintenance of an F' episome.

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Citations

Aug 27, 2016·Frontiers in Molecular Biosciences·Katarzyna E WegrzynIgor Konieczny
Mar 8, 2017·Biomolecular NMR Assignments·Esmael M AlyamiDmitry M Korzhnev
Apr 30, 2019·PLoS Computational Biology·Qiwen SunMoxun Tang
May 26, 2020·Critical Reviews in Biochemistry and Molecular Biology·Huidong Zhang
Apr 4, 2018·Proceedings of the National Academy of Sciences of the United States of America·Katarzyna H MaslowskaRoel M Schaaper
May 29, 2020·Molecular Cell·Enrico MonachinoAntoine M van Oijen
May 23, 2021·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Yu ZhaoQingtian Meng
Jun 2, 2021·EcoSal Plus·Maxime LerouxRodrigo Reyes-Lamothe
Sep 8, 2017·Chemical Research in Toxicology·Michael A TrakselisAurea M Chu
Sep 21, 2021·Journal of Bacteriology·Caleb HomiskiMark D Sutton

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