Heterologous expression of Na+/H+ antiporter gene (CvNHA1) from salt-tolerant yeast Candida versatilis in Saccharomyces cerevisiae Na+-transporter deficient mutants.
A Na(+)/H(+) antiporter gene (CvNHA1) was cloned from the salt-tolerant yeast Candida versatilis. CvNHA1 encodes an antiporter with a typical yeast plasma membrane Na(+)/H(+) antiporter structure. Transcription of CvNHA1 in C. versatilis cells was dependent on the salinity of the culture. When CvNHA1 was expressed in salt-sensitive Saccharomyces cerevisiae cells, increased salt-tolerance was observed, indicating that Cvnha1p possesses an Na(+)/H(+) antiporter function, because the increased salt-tolerance was dependent on the extracellular pH. It appears that Cvnha1p mediates only the transport of Na(+). In an S. cerevisiae transformant harboring a CvNHA1-EGFP fusion plasmid in which the greater part of the C-terminal hydrophilic region of Cvnha1p was deleted by fusion with enhanced green fluorescent protein (EGFP), the Cvnha1-EGFP fusion protein was localized mainly in the plasma membrane, and the NaCl-tolerance of this transformant was greater than that of a strain harboring the entire CvNHA1 gene.
Watanabe
Yasuo
Y
Department of Biological Resources, Faculty of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, Ehime 790-8566, Japan. watanabe@agr.ehime-u.ac.jp
Akita
Hidenori
H
Higuchi
Yuka
Y
Tsujimatsu
Rie
R
Kaneta
Tsuyoshi
T
Tamai
Youichi
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D002175:Candida / Q000187:drug effects / Q000235:genetics*; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D008969:Molecular Sequence Data; D009154:Mutation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D012492:Salts / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology; D017923:Sodium-Hydrogen Exchangers / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
D012492:Salts; D017923:Sodium-Hydrogen Exchangers; D012965:Sodium Chloride