title: Use of the cysteine-repressible HpMET3 promoter as a novel tool to regulate gene expression in Hansenula polymorpha.
abstract: OBJECTIVE
  The promoter of HpMET3, encoding an ATP sulfurylase, was evaluated for its potential as a repressible promoter to downregulate the expression of target genes in the thermotolerant, methylotrophic yeast Hansenula polymorpha.
  
  
  RESULTS
  The expression of lacZ under the control of the 0.6 kb HpMET3 promoter was efficiently downregulated by cysteine, but not by methionine or sulfate. The HpMET3 promoter was used to generate a conditional mutant of the HpPMT2 gene encoding an O-mannosyltransferase, which is involved in post-translational protein modification. The addition of 0.5 mM cysteine adversely affected the growth of the conditional HpMET3(p)-Hppmt2 mutant strain by downregulating transcription of HpPMT2 to approx. 40 % of the normal levels, indicating that the HpPMT2 gene is essential for cell viability. However, the HpMET3 promoter was neither induced nor repressed in the heterologous host Saccharomyces cerevisiae.
  
  
  CONCLUSIONS
  Our results reveal that the cysteine-repressible HpMET3 promoter is a useful tool that downregulates the expression of various genes in H. polymorpha.
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Korea.
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Yoo
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Korea.
authors.forename: Seung Yeon
authors.initials: SY
authors.lastname: Chung
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Korea.
authors.forename: Dong-Jik
authors.initials: DJ
authors.lastname: Lee
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Korea.
authors.forename: Hyunah
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Korea.
authors.forename: Seon Ah
authors.initials: SA
authors.lastname: Cheon
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, 156-756, Korea. hyunkang@cau.ac.kr.
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Kang
mesh_terms: D003545:Cysteine / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D008364:Mannosyltransferases / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013430:Sulfate Adenylyltransferase / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D008364:Mannosyltransferases; D013430:Sulfate Adenylyltransferase; D003545:Cysteine
keywords: ATP sulfurylase; Cysteine; Hansenula polymorpha; MET3; O-Mannosyltransferase; Promoter; Regulation