Characterization and regulation of Schizosaccharomyces pombe gene encoding thioredoxin

Young Wook Cho, Youn Hee Shin, Youn Tae Kim, HONGGYUM KIM, Yoon Jong Lee, Eun Hee Park, James A. Fuchs, Chang Jin Lim

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

A cDNA coding thioredoxin (TRX) was isolated from a cDNA library of Schizosaccharomyces pombe by colony hybridization. The 438 bp EcoRI fragment, which was detected by Southern hybridization, reveals an open reading frame which encodes a protein of 103 amino acids. The genomic DNA encoding TRX was also isolated from S. pombe chromosomal DNA using PCR. The cloned sequence contains 1795 bp and encodes a protein of 103 amino acids. However, the C-terminal region obtained from the cDNA clone is -Val-Arg-Leu-Asn-Arg-Ser-Leu, whereas the C-terminal region deduced from the genomic DNA appears to contain -Ala-Ser-Ile-Lys-Ala-Asn-Leu. This indicates that S. pombe cells contain two kinds of TRX genes which have dissimilar amino acid sequences only at the C-terminal regions. The heterologous TRX 1C produced from the cDNA clone could be used as a subunit of T7 DNA polymerase, while the TRX 1G from the genomic DNA could not. The upstream sequence and the region encoding the N-terminal 18 amino acids of the genomic DNA were fused into the promoterless β-galactosidase gene of the shuttle vector YEp357 to generate the fusion plasmid pYKT24. Synthesis of β-galactosidase from the fusion plasmid was found to be enhanced by hydrogen peroxide, menadione and aluminum chloride. It indicates that the expression of the cloned TRX gene is induced by oxidative stress.

Original languageEnglish (US)
Pages (from-to)194-199
Number of pages6
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1518
Issue number1-2
DOIs
StatePublished - Mar 19 2001

Bibliographical note

Funding Information:
This work was supported by a grant (No. 2000-1-20900-004-3) from the Basic Research Program of the Korea Science and Engineering Foundation.

Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.

Keywords

  • Fission yeast
  • Genomic DNA
  • Regulation
  • Schizosaccharomyces pombe
  • Thioredoxin
  • cDNA

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