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PMID: 10572171 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The top3(+) gene is essential in Schizosaccharomyces pombe and the lethality associated with its loss is caused by Rad12 helicase activity.

Nucleic acids research ·Vol. 27 ·No. 24 ·1999-12-15 ·Pages 4715-24

Maftahi M, Han CS, Langston LD, Hope JC, Zigouras N, Freyer GA

Abstract

The topoisomerase III gene ( top3 (+)) from Schizosaccharomyces pombe was isolated and a targeted gene disruption ( top3 :: kan (R)) was used to make a diploid strain heterozygous for top3 (+). The diploid was sporulated and the top3 :: kan (R)spores went through four to eight cell divisions before arresting as elongated, predominantly binucleated cells with incompletely segregated chromosomes. This demonstrates that top3 (+)is essential for vegetative growth in fission yeast. The aberrant chromosomal segregation seen in top3 :: kan (R)cells is unlike the 'cut' phenotype seen in mitosis-defective mutants and so we refer to this phenotype as 'torn'. A deletion mutant, rad12-hd ( rad12 is a homolog of Saccharomyces cerevisiae SGS1), partially suppressed the lethality of top3 mutants. A point mutant, rad12-K547I, which presumably eliminates helicase activity, also suppresses the lethality of top3 mutants, demonstrating that the lethality seen in top3 (-)cells is most likely caused by the helicase activity of Rad12. This double mutant grows very slowly and has much lower viability compared to rad12-hd top3 :: kan (R)cells, implying that the helicase activity of Rad12 is not the only cause of top3 (-)lethality. The low viability of rad12 (-) top3 (-)mutants compared with rad12 single mutants suggests that Top3 also functions independently of Rad12.

MeSH Terms
Amino Acid Sequence Amino Acid Substitution Cell Cycle Proteins/metabolism Cell Division Cloning, Molecular DNA Helicases/metabolism DNA Topoisomerases, Type I/chemistry,genetics,metabolism Diploidy Gene Deletion Genes, Essential Genes, Fungal Genes, Lethal Genotype Heterozygote Humans Mitosis Molecular Sequence Data Mutagenesis, Site-Directed Phenotype Recombinant Proteins/chemistry,metabolism Saccharomyces cerevisiae/enzymology,genetics Schizosaccharomyces/enzymology,genetics,physiology Schizosaccharomyces pombe Proteins Sequence Alignment Sequence Homology, Amino Acid Spores, Fungal
Chemicals
Cell Cycle Proteins Recombinant Proteins Schizosaccharomyces pombe Proteins DNA Helicases Rqh1 protein, S pombe DNA Topoisomerases, Type I
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Maftahi M
Division of Environmental Sciences, Joseph Mailman School of Public Health, New York, NY 10032, USA.
Han C S
Langston L D
Hope J C
Zigouras N
Freyer G A
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
Published
1999-12-15
Pages
4715-24
Language
English
Region
England
NLM ID
0411011
PMCID
PMC148771
Subset
IM
Grants
NCI NIH HHS · CA72647 · United States
NIEHS NIH HHS · ES-07940 · United States
Databases
GENBANK
AF126287
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