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Status |
Public on Feb 27, 2018 |
Title |
Evidence that DNA polymerase δ contributes to initiation of leading strand DNA replication in Saccharomyces cerevisiae |
Organism |
Saccharomyces cerevisiae |
Experiment type |
Other
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Summary |
To investigate nuclear DNA replication enzymology in vivo, we have studied Saccharomyces cerevisiae strains containing a pol2-16 mutation that inactivates the catalytic activities of DNA polymerase δ (Pol δ). Although pol2-16 mutants survive, their spore colonies are very tiny, with increased doubling time, larger than normal cells, aberrant nuclei, and rapid suppressor mutation accumulation. These phenotypes reveal a severe growth defect that is distinct from that of strains that lack Pol δ proofreading (pol2-4), consistent with the idea that Pol δ is the major leading strand replicase. Ribonucleotides are also incorporated into the pol2-16 genome in patterns consistent with leading strand replication by Pol δ when Pol δ is absent. More importantly, ribonucleotide distributions at replication origins suggest that in strains encoding all three replicases, Pol δ contributes to initiation of leading-strand replication. We describe two possible models.
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Overall design |
Examination of location of ribonucleotides in multiple yeast strains bearing different DNA polymerase variants, either proficient or deficient in rnh201.
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Contributor(s) |
Garbacz MA, Lujan SA, Burkholder AB, Cox PB, Wu Q, Zhou ZX, Haber JE, Kunkel TA |
Citation(s) |
29487291, 34019669 |
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Submission date |
Jul 20, 2017 |
Last update date |
Jul 25, 2021 |
Contact name |
Marta Anna Garbacz |
E-mail(s) |
garbaczmaw@gmail.com
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Phone |
9195410268
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Organization name |
NIEHS/NIH
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Department |
GISBL
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Lab |
DNA Replication Fidelity Lab
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Street address |
111 T.W. alexander Dr.
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City |
Research Triangle Park |
State/province |
NC |
ZIP/Postal code |
27713 |
Country |
USA |
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Platforms (1) |
GPL17342 |
Illumina HiSeq 2500 (Saccharomyces cerevisiae) |
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Samples (28)
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Relations |
BioProject |
PRJNA395222 |
SRA |
SRP113233 |
Supplementary file |
Size |
Download |
File type/resource |
GSE101698_RAW.tar |
396.0 Mb |
(http)(custom) |
TAR (of BW) |
GSE101698_RNR1.fa.gz |
3.5 Mb |
(ftp)(http) |
FA |
SRA Run Selector |
Raw data are available in SRA |
Processed data provided as supplementary file |
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