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DNA polymerase e leading strand signature mutations result from defects in its proofreading activity.

Authors :
Johnson, Robert E.
Prakash, Louise
Prakash, Satya
Source :
Journal of Biological Chemistry. Jul2023, Vol. 299 Issue 7, p1-8. 8p.
Publication Year :
2023

Abstract

The evidence that purified pol2-M644G DNA polymerase (Pol)e exhibits a highly elevated bias for forming T:dTTP mispairs over A:dATP mispairs and that yeast cells harboring this Pole mutation accumulate A > T signature mutations in the leading strand have been used to assign a role for Pole in replicating the leading strand. Here, we determine whether A > T signature mutations result from defects in Pole proofreading activity by analyzing their rate in Pole proofreading defective pol2-4 and pol2-M644G cells. Since purified pol2-4 Pole exhibits no bias for T:dTTP mispair formation, A > T mutations are expected to occur at a much lower rate in pol2-4 than in pol2-M644G cells if Pole replicated the leading strand. Instead, we find that the rate of A > T signature mutations are as highly elevated in pol2-4 cells as in pol2-M644G cells; furthermore, the highly elevated rate of A > T signature mutations is severely curtailed in the absence of PCNA ubiquitination or Pol? in both the pol2-M644G and pol2-4 strains. Altogether, our evidence supports the conclusion that the leading strand A > T signature mutations derive from defects in Pole proofreading activity and not from the role of Pole as a leading strand replicase, and it conforms with the genetic evidence for a major role of Pold in replication of both the DNA strands. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00219258
Volume :
299
Issue :
7
Database :
Academic Search Index
Journal :
Journal of Biological Chemistry
Publication Type :
Academic Journal
Accession number :
169843241
Full Text :
https://doi.org/10.1016/j.jbc.2023.104913