A role for DNA primase in coupling DNA replication to DNA damage response.
The temperature-sensitive yeast DNA primase mutant pri1-M4 fails to execute an early step of DNA replication and exhibits a dominant, allele-specific sensitivity to DNA-damaging agents. pri1-M4 is defective in slowing down the rate of S phase progression and partially delaying the G1-S transition in response to DNA damage. Conversely, the G2 DNA damage response and the S-M checkpoint coupling completion of DNA replication to mitosis are unaffected. The signal transduction pathway leading to Rad53p phosphorylation induced by DNA damage is proficient in pri1-M4, and cell cycle delay caused by Rad53p overexpression is counteracted by the pri1-M4 mutation. Altogether, our results suggest that DNA primase plays an essential role in a subset of the Rad53p-dependent checkpoint pathways controlling cell cycle progression in response to DNA damage.
Marini
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F
Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Italy.
Pellicioli
A
A
Paciotti
V
V
Lucchini
G
G
Plevani
P
P
Stern
D F
DF
Foiani
M
M
D015153:Blotting, Western; D002453:Cell Cycle / Q000235:genetics; D018797:Cell Cycle Proteins; D064447:Checkpoint Kinase 2; D004247:DNA / Q000096:biosynthesis; D004249:DNA Damage / Q000235:genetics*; D019915:DNA Primase; D004261:DNA Replication / Q000235:genetics*; D004795:Enzyme Stability / Q000235:genetics; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D007399:Interphase / Q000235:genetics; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008938:Mitosis / Q000235:genetics; D008954:Models, Biological; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D009153:Mutagens / Q000494:pharmacology; D009154:Mutation / Q000235:genetics; D010766:Phosphorylation; D011494:Protein Kinases; D017346:Protein Serine-Threonine Kinases; D012316:RNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D016196:S Phase / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013696:Temperature; D014466:Ultraviolet Rays / Q000009:adverse effects
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D009153:Mutagens; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D008741:Methyl Methanesulfonate; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; D017346:Protein Serine-Threonine Kinases; C067022:RAD53 protein, S cerevisiae; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases