The absence of ribonuclease H1 or H2 alters the sensitivity of Saccharomyces cerevisiae to hydroxyurea, caffeine and ethyl methanesulphonate: implications for roles of RNases H in DNA replication and repair.
BACKGROUND
RNA of RNA-DNA hybrids can be degraded by ribonucleases H present in all organisms including the eukaryote Saccharomyces cerevisiae. Determination of the number and roles of the RNases H in eukaryotes is quite feasible in S. cerevisiae.


RESULTS
Two S. cerevisiae RNases H, related to Escherichia coli RNase HI and HII, are not required for growth under normal conditions, yet, compared with wild-type cells, a double-deletion strain has an increased sensitivity to hydroxyurea (HU) and is hypersensitive to caffeine and ethyl methanesulphonate (EMS). In the absence of RNase H1, RNase H2 activity increases, and cells are sensitive to EMS but not HU and are more tolerant of caffeine; the latter requires RNase H2 activity. Cells missing only RNase H2 exhibit increased sensitive to HU and EMS but not caffeine


CONCLUSIONS
Mutant phenotypes infer that some RNA-DNA hybrids are recognized by both RNases H1 and H2, while other hybrids appear to be recognized only by RNase H2. Undegraded RNA-DNA hybrids have an effect when DNA synthesis is impaired, DNA damage occurs or the cell cycle is perturbed by exposure to caffeine suggesting a role in DNA replication/repair that can be either beneficial or detrimental to cell viability.
Arudchandran
A
A
Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892-2790, USA.
Cerritelli
S
S
Narimatsu
S
S
Itaya
M
M
Shin
D Y
DY
Shimada
Y
Y
Crouch
R J
RJ
D002110:Caffeine / Q000494:pharmacology*; D002453:Cell Cycle; D002467:Cell Nucleus / Q000201:enzymology; D004260:DNA Repair; D004261:DNA Replication; D004271:DNA, Fungal / Q000096:biosynthesis / Q000378:metabolism; D005020:Ethyl Methanesulfonate / Q000494:pharmacology*; D017353:Gene Deletion; D005800:Genes, Fungal; D006918:Hydroxyurea / Q000494:pharmacology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010641:Phenotype; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics
D004271:DNA, Fungal; D019384:Nucleic Acid Synthesis Inhibitors; D011993:Recombinant Fusion Proteins; D002110:Caffeine; D005020:Ethyl Methanesulfonate; C091139:ribonuclease HII; D016914:Ribonuclease H; C078856:ribonuclease HI; D006918:Hydroxyurea