Different domains of Sgs1 are required for mitotic and meiotic functions.
The SGS1 of Saccharomyces cerevisiae is a homologue for human Bloom's syndrome, Werner's syndrome, and Rothmund-Thomson's syndrome causative genes. Disruptants of SGS1 show high sensitivity to methyl methanesulfonate (MMS) and hydroxyurea, and hyper recombination phenotypes including interchromosomal homologous recombination in mitotic growth. In addition, sgs1 disruptants show poor sporulation and a reduced level of meiotic recombination as assayed by return-to-growth. We examined domains of Sgs1 required for mitotic and meiotic functions of Sgs1 by transfecting variously mutated SGS1 into sgs1 disruptants. The N-terminal 1-401 amino acid region was required for complementation of MMS sensitivity and suppression of hyper heteroallelic recombinations of sgs1 disruptants in mitotic growth and for complementation of poor sporulation and of reduced meiotic recombination. Although the N-terminal 1-125 amino acid region was absolutely required for the complementation of MMS sensitivity and suppression of hyper heteroallelic recombinations in mitotic growth, it was dispensable for the meiotic functions. In contrast, the highly acidic region (400-596 amino acid) was dispensable for the mitotic functions but a deletion of this region affected the meiotic functions. The C-terminal 1271-1350 amino acid region containing a HRDC (helicase and RNaseD C-terminal) domain was dispensable for the mitotic and meiotic functions. Although DNA helicase activity of Sgs1 was not required for Sgs1 to complement the meiotic functions, a deletion of helicase motifs III-IV (842-1046 amino acid) abolished the complementing activity of Sgs1, indicating that a structurally intact helicase domain is necessary for Sgs1 to fulfill its meiotic functions.
Division of Pharmacology, Biological Safety Research Center, National Institute of Health Sciences, Setagaya-ku, Tokyo 158-8501, Japan.
A
A
Miyajima
M
M
Seki
F
F
Onoda
A
A
Ui
Y
Y
Satoh
Y
Y
Ohno
T
T
Enomoto
D000483:Alleles; D018797:Cell Cycle Proteins / Q000378:metabolism; D004265:DNA Helicases / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005656:Fungal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D005838:Genotype; D018398:Homeodomain Proteins; D006918:Hydroxyurea / Q000494:pharmacology*; D008540:Meiosis; D008741:Methyl Methanesulfonate / Q000494:pharmacology*; D008938:Mitosis; D016297:Mutagenesis, Site-Directed; D009153:Mutagens / Q000494:pharmacology; D020125:Mutation, Missense; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D017386:Sequence Homology, Amino Acid; D012854:Sister Chromatid Exchange / Q000235:genetics*; D013172:Spores, Fungal / Q000187:drug effects* / Q000254:growth & development; D014162:Transfection
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D018398:Homeodomain Proteins; D009153:Mutagens; D008741:Methyl Methanesulfonate; D004265:DNA Helicases; D006918:Hydroxyurea