ntf1+ encodes a 6-cysteine zinc finger-containing transcription factor that regulates the nmt1 promoter in fission yeast.
The nmt1+ gene of the fission yeast Schizosaccharomyces pombe is subject to transcriptional repression mediated by thiamine. The promoter of nmt1+ has been used to construct a series of vectors that are now commonly used for the regulated expression of genes in S. pombe. In this report, we described ntf1+, a gene involved in regulating nmt1+ expression. The ntf1+ gene was cloned in a high copy suppressor screen that utilized a construct, nmt1:wee1+, in which expression of the mitotic inhibitor Wee1 tyrosine kinase was placed under the control of the nmt1 promoter. ntf1+ encodes a 706-amino acid protein that shares substantial homology with a family of Cys6 zinc finger-containing transcription factors typified by GAL4 from Saccharomyces cerevisiae. Increased gene dosage of ntf1+ greatly increases both the repressed and derepressed activity of the nmt1 promoter. Cells having a disrupted version of ntf1+ are viable thiamine prototrophs, but basal expression from the nmt1 promoter is greatly reduced. These data demonstrate that Ntf1 plays an important role in regulating nmt1 expression.
Tang
C S
CS
Department of Molecular Biology, Scripps Research Institute, La Jolla, California 92037.
Bueno
A
A
Russell
P
P
D000595:Amino Acid Sequence; D001483:Base Sequence; D003545:Cysteine; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D012568:Schizosaccharomyces / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D016335:Zinc Fingers / Q000235:genetics*
D004268:DNA-Binding Proteins; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C086956:thi1 protein, S pombe; D003545:Cysteine