Identification of the Cadmium-Inducible Hansenula polymorpha SEO1 Gene Promoter by Transcriptome Analysis and Its Application to Whole-Cell Heavy-Metal Detection Systems

ABSTRACT The genomewide gene expression profiling of the methylotrophic yeast Hansenula polymorpha exposed to cadmium (Cd) allowed us to identify novel genes responsive to Cd treatment. To select genes whose promoters can be useful for construction of a cellular Cd biosensor, we further analyzed a set of H. polymorpha genes that exhibited >6-fold induction upon treatment with 300 μM Cd for 2 h. The putative promoters, about 1,000-bp upstream fragments, of these genes were fused with the yeast-enhanced green fluorescence protein (GFP) gene. The resultant reporter cassettes were introduced into H. polymorpha to evaluate promoter strength and specificity. The promoter derived from the H. polymorpha SEO1 gene (HpSEO1) was shown to drive most strongly the expression of GFP upon Cd treatment among the tested promoters. The Cd-inducible activity was retained in the 500-bp deletion fragment of the HpSEO1 promoter but was abolished in the further truncated 250-bp fragment. The 500-bp HpSEO1 promoter directed specific expression of GFP upon exposure to Cd in a dose-dependent manner, with Cd detection ranging from 1 to 900 μM. Comparative analysis of the Saccharomyces cerevisiae SEO1 (ScSEO1) promoter revealed that the ScSEO1 promoter has a broader specificity for heavy metals and is responsive to arsenic and mercury in addition to Cd. Our data demonstrate the potential use of the HpSEO1 promoter as a bioelement in whole-cell biosensors to monitor heavy metal contamination, particularly Cd.

[1]  A. Myers,et al.  Yeast/E. coli shuttle vectors with multiple unique restriction sites , 1986, Yeast.

[2]  D. E. Griffiths,et al.  DMSO-enhanced whole cell yeast transformation. , 1991, Nucleic acids research.

[3]  L. Zhu,et al.  Isolation of genomic DNAs from plants, fungi and bacteria using benzyl chloride. , 1993, Nucleic acids research.

[4]  C. S. Parker,et al.  Transcriptional activation mediated by the yeast AP-1 protein is required for normal cadmium tolerance. , 1994, The Journal of biological chemistry.

[5]  Y. Surdin-Kerjan,et al.  The study of methionine uptake in Saccharomyces cerevisiae reveals a new family of amino acid permeases. , 1996, Journal of molecular biology.

[6]  A. Marchler-Bauer,et al.  The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). , 1996, The EMBO journal.

[7]  Dominique Thomas,et al.  Assembly of a bZIP–bHLH transcription activation complex: formation of the yeast Cbf1–Met4–Met28 complex is regulated through Met28 stimulation of Cbf1 DNA binding , 1997, The EMBO journal.

[8]  Rupert De Wachter,et al.  Classification of all putative permeases and other membrane multispanners of the Major Facilitator Superfamily encoded by the complete genome of Saccharomyces cerevisiae , 1997, German Conference on Bioinformatics.

[9]  I. Mannazzu,et al.  The vanadate-tolerant yeast Hansenula polymorpha undergoes cellular reorganization during growth in, and recovery from, the presence of vanadate. , 1998, Microbiology.

[10]  M Virta,et al.  Luminescent bacterial sensor for cadmium and lead. , 1998, Biosensors & bioelectronics.

[11]  G. Church,et al.  Systematic determination of genetic network architecture , 1999, Nature Genetics.

[12]  B. Rosen,et al.  Pathways of As(III) detoxification in Saccharomyces cerevisiae. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[13]  J. Sohn,et al.  Proteolytic stability of recombinant human serum albumin secreted in the yeast Saccharomyces cerevisiae , 2000, Applied Microbiology and Biotechnology.

[14]  C. Martelet,et al.  Development of highly selective and stable potentiometric sensors for formaldehyde determination. , 2000, Biosensors & bioelectronics.

[15]  J. Mellor,et al.  Cadmium-inducible Expression of the Yeast GSH1 Gene Requires a Functional Sulfur-Amino Acid Regulatory Network* , 2000, The Journal of Biological Chemistry.

[16]  I. Mannazzu,et al.  Vanadate and copper induce overlapping oxidative stress responses in the vanadate-tolerant yeast Hansenula polymorpha. , 2000, Biochimica et biophysica acta.

[17]  M. Lehmann,et al.  Amperometric measurement of copper ions with a deputy substrate using a novel Saccharomyces cerevisiae sensor. , 2000, Biosensors & bioelectronics.

[18]  H. Iwahashi,et al.  Bioassay of cadmium using a DNA microarray: Genome‐wide expression patterns of Saccharomyces cerevisiae response to cadmium , 2001, Environmental toxicology and chemistry.

[19]  J. Sohn,et al.  Development of expression systems for the production of recombinant human serum albumin using the MOX promoter in Hansenula polymorpha DL-1. , 2001, Biotechnology and bioengineering.

[20]  D. Eide.  Functional genomics and metal metabolism , 2001, Genome Biology.

[21]  M. Toledano,et al.  A Proteome Analysis of the Cadmium Response in Saccharomyces cerevisiae * , 2001, The Journal of Biological Chemistry.

[22]  P. Wingfield.  Production of Recombinant Proteins , 2002 .

[23]  B. Rosen.  Transport and detoxification systems for transition metals, heavy metals and metalloids in eukaryotic and prokaryotic microbes. , 2002, Comparative biochemistry and physiology. Part A, Molecular & integrative physiology.

[24]  G. Gellissen.  Hansenula polymorpha: Biology and Applications , 2002 .

[25]  D. Jamieson,et al.  The adaptive response of Saccharomyces cerevisiae to mercury exposure , 2002, Yeast.

[26]  Krishnamurthy Natarajan,et al.  Gcn4p, a Master Regulator of Gene Expression, Is Controlled at Multiple Levels by Diverse Signals of Starvation and Stress , 2002, Eukaryotic Cell.

[27]  Michel Werner,et al.  Sulfur sparing in the yeast proteome in response to sulfur demand. , 2002, Molecular cell.

[28]  Y. Korpan,et al.  Metabolically engineered methylotrophic yeast cells and enzymes as sensor biorecognition elements. , 2002, FEMS yeast research.

[29]  Jürg Bähler,et al.  Whole-genome microarrays of fission yeast: characteristics, accuracy, reproducibility, and processing of array data , 2003, BMC Genomics.

[30]  S. Belkin.  Microbial whole-cell sensing systems of environmental pollutants. , 2003, Current opinion in microbiology.

[31]  C. Hollenberg,et al.  The Hansenula polymorpha (strain CBS4732) genome sequencing and analysis. , 2003, FEMS yeast research.

[32]  M. Lazard,et al.  Ycf1p-dependent Hg(II) detoxification in Saccharomyces cerevisiae. , 2003, European journal of biochemistry.

[33]  B. Birren,et al.  Sequencing and comparison of yeast species to identify genes and regulatory elements , 2003, Nature.

[34]  A. Brazma,et al.  Global transcriptional responses of fission yeast to environmental stress. , 2003, Molecular biology of the cell.

[35]  N. Verma,et al.  Biosensors for heavy metals , 2005, Biometals.

[36]  Man Bock Gu,et al.  Whole-cell-based biosensors for environmental biomonitoring and application. , 2004, Advances in biochemical engineering/biotechnology.

[37]  G. Gellissen,et al.  Fabrication of a Partial Genome Microarray of the Methylotrophic Yeast Hansenula polymorpha: Optimization and Evaluation of Transcript Profiling , 2004 .

[38]  S. Leonard,et al.  Cadmium inhibits the electron transfer chain and induces reactive oxygen species. , 2004, Free radical biology & medicine.

[39]  J. Mellor,et al.  Cbf1p Is Required for Chromatin Remodeling at Promoter-proximal CACGTG Motifs in Yeast* , 2004, Journal of Biological Chemistry.

[40]  K. Baronian,et al.  The use of yeast and moulds as sensing elements in biosensors. , 2004, Biosensors & bioelectronics.

[41]  Y. Shimma,et al.  Characterization of N-linked oligosaccharides assembled on secretory recombinant glucose oxidase and cell wall mannoproteins from the methylotrophic yeast Hansenula polymorpha. , 2003, Glycobiology.

[42]  S. Palecek,et al.  Saccharomyces cerevisiae JEN1 Promoter Activity Is Inversely Related to Concentration of Repressing Sugar , 2004, Applied and Environmental Microbiology.

[43]  L. A. Palomares,et al.  Production of recombinant proteins: challenges and solutions. , 2004, Methods in molecular biology.

[44]  R. Moreno-Sánchez,et al.  Sulfur assimilation and glutathione metabolism under cadmium stress in yeast, protists and plants. , 2005, FEMS microbiology reviews.

[45]  Man Bock Gu,et al.  Screening of target-specific stress-responsive genes for the development of cell-based biosensors using a DNA microarray. , 2005, Analytical chemistry.

[46]  Hwan-Gue Cho,et al.  An Open Source Microarray Data Analysis System with GUI: Quintet , 2005, RSFDGrC.

[47]  H. Kang,et al.  Accumulation of cadmium ions in the methylotrophic yeast Hansenula polymorpha , 2006, Biometals.

[48]  Teresa Lettieri,et al.  Recent Applications of DNA Microarray Technology to Toxicology and Ecotoxicology , 2005, Environmental health perspectives.

[49]  H. Nanjo,et al.  Naked-eye cadmium sensor: using chromoionophore arrays of Langmuir-Blodgett molecular assemblies. , 2007, Analytical chemistry.