scholarly journals Regulation of yeast DNA polymerase δ-mediated strand displacement synthesis by 5′-flaps

2015 ◽  
Vol 43 (8) ◽  
pp. 4179-4190 ◽  
Author(s):  
Katrina N. Koc ◽  
Joseph L. Stodola ◽  
Peter M. Burgers ◽  
Roberto Galletto
Keyword(s):  
Dna Polymerase ◽  
Strand Displacement ◽  
Dna Polymerase Δ

scholarly journals Pif1, RPA, and FEN1 modulate the ability of DNA polymerase δ to overcome protein barriers during DNA synthesis

2020 ◽  
Vol 295 (47) ◽  
pp. 15883-15891 ◽  
Author(s):  
Melanie A. Sparks ◽  
Peter M. Burgers ◽  
Roberto Galletto
Keyword(s):  
Transcription Factors ◽  
Dna Replication ◽  
Dna Binding ◽  
Dna Synthesis ◽  
Dna Polymerase ◽  
Strand Displacement ◽  
Chromosomal Dna ◽  
Dna Polymerase Δ ◽  
Lagging Strand ◽  
Stimulation Of

Successful DNA replication requires carefully regulated mechanisms to overcome numerous obstacles that naturally occur throughout chromosomal DNA. Scattered across the genome are tightly bound proteins, such as transcription factors and nucleosomes, that are necessary for cell function, but that also have the potential to impede timely DNA replication. Using biochemically reconstituted systems, we show that two transcription factors, yeast Reb1 and Tbf1, and a tightly positioned nucleosome, are strong blocks to the strand displacement DNA synthesis activity of DNA polymerase δ. Although the block imparted by Tbf1 can be overcome by the DNA-binding activity of the single-stranded DNA-binding protein RPA, efficient DNA replication through either a Reb1 or a nucleosome block occurs only in the presence of the 5'-3' DNA helicase Pif1. The Pif1-dependent stimulation of DNA synthesis across strong protein barriers may be beneficial during break-induced replication where barriers are expected to pose a problem to efficient DNA bubble migration. However, in the context of lagging strand DNA synthesis, the efficient disruption of a nucleosome barrier by Pif1 could lead to the futile re-replication of newly synthetized DNA. In the presence of FEN1 endonuclease, the major driver of nick translation during lagging strand replication, Pif1-dependent stimulation of DNA synthesis through a nucleosome or Reb1 barrier is prevented. By cleaving the short 5' tails generated during strand displacement, FEN1 eliminates the entry point for Pif1. We propose that this activity would protect the cell from potential DNA re-replication caused by unwarranted Pif1 interference during lagging strand replication.

scholarly journals Pif1-family helicases cooperate to suppress widespread replication fork arrest at tRNA genes

2016 ◽  
Author(s):  
Joseph S. Osmundson ◽  
Jayashree Kumar ◽  
Rani Yeung ◽  
Duncan J. Smith
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Damage ◽  
Dna Polymerase ◽  
Replication Fork ◽  
Trna Genes ◽  
Strand Displacement ◽  
Dna Polymerase Δ ◽  
Replication Fork Arrest ◽  
Nuclear Processes ◽  
Lagging Strand

ABSTRACTSaccharomyces cerevisiaeencodes two distinct Pif1-family helicases – Pif1 and Rrm3 – which have been reported to play distinct roles in numerous nuclear processes. Here, we systematically characterize the roles of Pif1 helicases in replisome progression and lagging-strand synthesis inS. cerevisiae. We demonstrate that either Pif1 or Rrm3 redundantly stimulate strand-displacement by DNA polymerase δ during lagging-strand synthesis. By analyzing replisome mobility inpif1andrrm3mutants, we show that Rrm3, with a partially redundant contribution from Pif1, suppresses widespread terminal arrest of the replisome at tRNA genes. Although both head-on and codirectional collisions induce replication fork arrest at tRNA genes, head-on collisions arrest a higher proportion of replisomes; consistent with this observation, we find that head-on collisions between tRNA transcription and replisome progression are under-represented in theS. cerevisiaegenome. Further, we demonstrate that tRNA-mediated arrest is R-loop independent, and propose that replisome arrest and DNA damage are mechanistically separable.

scholarly journals Pif1 removes a Rap1-dependent barrier to the strand displacement activity of DNA polymerase δ

2016 ◽  
Vol 44 (8) ◽  
pp. 3811-3819 ◽  
Author(s):  
Katrina N. Koc ◽  
Saurabh P. Singh ◽  
Joseph L. Stodola ◽  
Peter M. Burgers ◽  
Roberto Galletto
Keyword(s):  
Dna Polymerase ◽  
Strand Displacement ◽  
Displacement Activity ◽  
Dna Polymerase Δ

scholarly journals Underappreciated Roles of DNA Polymerase δ in Replication Stress Survival

2021 ◽  
Vol 37 (5) ◽  
pp. 476-487 ◽  
Author(s):  
Jeannette Fuchs ◽  
Anais Cheblal ◽  
Susan M. Gasser
Keyword(s):  
Dna Polymerase ◽  
Replication Stress ◽  
Dna Polymerase Δ ◽  
Stress Survival

scholarly journals DNA polymerase δ stalls on telomeric lagging strand templates independently from G-quadruplex formation

2013 ◽  
Vol 41 (22) ◽  
pp. 10323-10333 ◽  
Author(s):  
Justin D. Lormand ◽  
Noah Buncher ◽  
Connor T. Murphy ◽  
Parminder Kaur ◽  
Marietta Y. Lee ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerase Δ ◽  
G Quadruplex ◽  
Quadruplex Formation ◽  
Lagging Strand

scholarly journals A Mutation of the Yeast Gene Encoding PCNA Destabilizes Both Microsatellite and Minisatellite DNA Sequences

Genetics ◽  
1999 ◽  
Vol 151 (2) ◽  
pp. 511-519 ◽  
Author(s):  
Robert J Kokoska ◽  
Lela Stefanovic ◽  
Andrew B Buermeyer ◽  
R Michael Liskay ◽  
Thomas D Petes
Keyword(s):  
Dna Polymerase ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Repeat Unit ◽  
Nuclear Antigen ◽  
Temperature Sensitive ◽  
Dinucleotide Repeats ◽  
Yeast Saccharomyces Cerevisiae ◽  
Dna Polymerase Δ ◽  
Repeat Units

AbstractThe POL30 gene of the yeast Saccharomyces cerevisiae encodes the proliferating cell nuclear antigen (PCNA), a protein required for processive DNA synthesis by DNA polymerase δ and ϵ. We examined the effects of the pol30-52 mutation on the stability of microsatellite (1- to 8-bp repeat units) and minisatellite (20-bp repeat units) DNA sequences. It had previously been shown that this mutation destabilizes dinucleotide repeats 150-fold and that this effect is primarily due to defects in DNA mismatch repair. From our analysis of the effects of pol30-52 on classes of repetitive DNA with longer repeat unit lengths, we conclude that this mutation may also elevate the rate of DNA polymerase slippage. The effect of pol30-52 on tracts of repetitive DNA with large repeat unit lengths was similar, but not identical, to that observed previously for pol3-t, a temperature-sensitive mutation affecting DNA polymerase δ. Strains with both pol30-52 and pol3-t mutations grew extremely slowly and had minisatellite mutation rates considerably greater than those observed in either single mutant strain.

Sign in / Sign up

Export Citation Format

Share Document