New insights into galactose metabolism by Schizosaccharomyces pombe: Isolation and characterization of a galactose-assimilating mutant

Tomohiko Matsuzawa, Yasuko Fujita, Naotaka Tanaka, Hideki Tohda, Akiko Itadani, Kaoru Takegawa

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The fission yeast Schizosaccharomyces pombe cannot use galactose as a carbon or energy source, and little is known about galactose metabolism in this species. Here we report isolation of a galactose-assimilating mutant that grows on a medium containing galactose as a sole carbon source through use of a proofreading-deficient DNA polymerase δ variant encoded by cdc6-1. Based on comparative analysis of gene expression profiles in the wild-type and the mutant (FG2-8), we found that SPBPB2B2.10c (gal7+), SPBPB2B2.12c (gal10+) and SPBPB2B2.13 (gal1+), homologous to Saccharomyces cerevisiae GAL7, GAL10 and GAL1, respectively, and SPBPB2B2.08, SPBPB2B2.09c, and SPBPB2B2.11 that localize close to the gal genes, were highly expressed and dramatically induced by addition of galactose. The gal7δ strain, carrying an integrated ura4+ marker at the gal7+ locus, grew on 5-fluoroorotic acid (5-FOA)-containing medium. In contrast, the FG2-8 gal7δ strain could not grow on 5-FOA medium. In addition, expression of gal7+, SPBPB2B2.13, gal10+ and gal1+ genes increased in the wild-type strain when carried on a vector, and these transformants grew on galactose medium. We suggest that gal7+, gal10+, and gal1+ are localized close to a chromosomal terminal repressed by gene silencing in S. pombe. In contrast, gene silencing was defective in the FG2-8 strain making galactose assimilation possible.

Original languageEnglish
Pages (from-to)158-166
Number of pages9
JournalJournal of Bioscience and Bioengineering
Volume111
Issue number2
DOIs
Publication statusPublished - Feb 1 2011

Fingerprint

Schizosaccharomyces
Galactose
Metabolism
Genes
Yeast
Gene Silencing
Carbon
Acids
Gene expression
DNA
DNA-Directed DNA Polymerase
Transcriptome
Saccharomyces cerevisiae

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

New insights into galactose metabolism by Schizosaccharomyces pombe : Isolation and characterization of a galactose-assimilating mutant. / Matsuzawa, Tomohiko; Fujita, Yasuko; Tanaka, Naotaka; Tohda, Hideki; Itadani, Akiko; Takegawa, Kaoru.

In: Journal of Bioscience and Bioengineering, Vol. 111, No. 2, 01.02.2011, p. 158-166.

Research output: Contribution to journalArticle

Matsuzawa, Tomohiko ; Fujita, Yasuko ; Tanaka, Naotaka ; Tohda, Hideki ; Itadani, Akiko ; Takegawa, Kaoru. / New insights into galactose metabolism by Schizosaccharomyces pombe : Isolation and characterization of a galactose-assimilating mutant. In: Journal of Bioscience and Bioengineering. 2011 ; Vol. 111, No. 2. pp. 158-166.
@article{15988377722d48609dc07ff8e1fc8e50,
title = "New insights into galactose metabolism by Schizosaccharomyces pombe: Isolation and characterization of a galactose-assimilating mutant",
abstract = "The fission yeast Schizosaccharomyces pombe cannot use galactose as a carbon or energy source, and little is known about galactose metabolism in this species. Here we report isolation of a galactose-assimilating mutant that grows on a medium containing galactose as a sole carbon source through use of a proofreading-deficient DNA polymerase δ variant encoded by cdc6-1. Based on comparative analysis of gene expression profiles in the wild-type and the mutant (FG2-8), we found that SPBPB2B2.10c (gal7+), SPBPB2B2.12c (gal10+) and SPBPB2B2.13 (gal1+), homologous to Saccharomyces cerevisiae GAL7, GAL10 and GAL1, respectively, and SPBPB2B2.08, SPBPB2B2.09c, and SPBPB2B2.11 that localize close to the gal genes, were highly expressed and dramatically induced by addition of galactose. The gal7δ strain, carrying an integrated ura4+ marker at the gal7+ locus, grew on 5-fluoroorotic acid (5-FOA)-containing medium. In contrast, the FG2-8 gal7δ strain could not grow on 5-FOA medium. In addition, expression of gal7+, SPBPB2B2.13, gal10+ and gal1+ genes increased in the wild-type strain when carried on a vector, and these transformants grew on galactose medium. We suggest that gal7+, gal10+, and gal1+ are localized close to a chromosomal terminal repressed by gene silencing in S. pombe. In contrast, gene silencing was defective in the FG2-8 strain making galactose assimilation possible.",
author = "Tomohiko Matsuzawa and Yasuko Fujita and Naotaka Tanaka and Hideki Tohda and Akiko Itadani and Kaoru Takegawa",
year = "2011",
month = "2",
day = "1",
doi = "10.1016/j.jbiosc.2010.10.007",
language = "English",
volume = "111",
pages = "158--166",
journal = "Journal of Bioscience and Bioengineering",
issn = "1389-1723",
publisher = "The Society for Biotechnology, Japan",
number = "2",

}

TY - JOUR

T1 - New insights into galactose metabolism by Schizosaccharomyces pombe

T2 - Isolation and characterization of a galactose-assimilating mutant

AU - Matsuzawa, Tomohiko

AU - Fujita, Yasuko

AU - Tanaka, Naotaka

AU - Tohda, Hideki

AU - Itadani, Akiko

AU - Takegawa, Kaoru

PY - 2011/2/1

Y1 - 2011/2/1

N2 - The fission yeast Schizosaccharomyces pombe cannot use galactose as a carbon or energy source, and little is known about galactose metabolism in this species. Here we report isolation of a galactose-assimilating mutant that grows on a medium containing galactose as a sole carbon source through use of a proofreading-deficient DNA polymerase δ variant encoded by cdc6-1. Based on comparative analysis of gene expression profiles in the wild-type and the mutant (FG2-8), we found that SPBPB2B2.10c (gal7+), SPBPB2B2.12c (gal10+) and SPBPB2B2.13 (gal1+), homologous to Saccharomyces cerevisiae GAL7, GAL10 and GAL1, respectively, and SPBPB2B2.08, SPBPB2B2.09c, and SPBPB2B2.11 that localize close to the gal genes, were highly expressed and dramatically induced by addition of galactose. The gal7δ strain, carrying an integrated ura4+ marker at the gal7+ locus, grew on 5-fluoroorotic acid (5-FOA)-containing medium. In contrast, the FG2-8 gal7δ strain could not grow on 5-FOA medium. In addition, expression of gal7+, SPBPB2B2.13, gal10+ and gal1+ genes increased in the wild-type strain when carried on a vector, and these transformants grew on galactose medium. We suggest that gal7+, gal10+, and gal1+ are localized close to a chromosomal terminal repressed by gene silencing in S. pombe. In contrast, gene silencing was defective in the FG2-8 strain making galactose assimilation possible.

AB - The fission yeast Schizosaccharomyces pombe cannot use galactose as a carbon or energy source, and little is known about galactose metabolism in this species. Here we report isolation of a galactose-assimilating mutant that grows on a medium containing galactose as a sole carbon source through use of a proofreading-deficient DNA polymerase δ variant encoded by cdc6-1. Based on comparative analysis of gene expression profiles in the wild-type and the mutant (FG2-8), we found that SPBPB2B2.10c (gal7+), SPBPB2B2.12c (gal10+) and SPBPB2B2.13 (gal1+), homologous to Saccharomyces cerevisiae GAL7, GAL10 and GAL1, respectively, and SPBPB2B2.08, SPBPB2B2.09c, and SPBPB2B2.11 that localize close to the gal genes, were highly expressed and dramatically induced by addition of galactose. The gal7δ strain, carrying an integrated ura4+ marker at the gal7+ locus, grew on 5-fluoroorotic acid (5-FOA)-containing medium. In contrast, the FG2-8 gal7δ strain could not grow on 5-FOA medium. In addition, expression of gal7+, SPBPB2B2.13, gal10+ and gal1+ genes increased in the wild-type strain when carried on a vector, and these transformants grew on galactose medium. We suggest that gal7+, gal10+, and gal1+ are localized close to a chromosomal terminal repressed by gene silencing in S. pombe. In contrast, gene silencing was defective in the FG2-8 strain making galactose assimilation possible.

UR - http://www.scopus.com/inward/record.url?scp=78951493734&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78951493734&partnerID=8YFLogxK

U2 - 10.1016/j.jbiosc.2010.10.007

DO - 10.1016/j.jbiosc.2010.10.007

M3 - Article

C2 - 21075050

AN - SCOPUS:78951493734

VL - 111

SP - 158

EP - 166

JO - Journal of Bioscience and Bioengineering

JF - Journal of Bioscience and Bioengineering

SN - 1389-1723

IS - 2

ER -