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A new nuclear suppressor system for a mitochondrial RNA polymerase mutant identifies an unusual zinc‐finger protein and a polyglutamine domain protein in Saccharomyces cerevisiae
Author(s) -
Bröhl Stefanie,
Lisowsky Thomas,
Riemen Gudula,
Michaelis Georg
Publication year - 1994
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320100604
Subject(s) - biology , zinc finger , gene , mutant , microbiology and biotechnology , ring finger domain , genetics , protein subunit , transcription factor
A yeast strain with a point mutation in the nuclear gene for the core subunit of mitochondrial RNA polymerase was used to isolate new extragenic suppressors. Spontaneously occurring phenotypical revertants were analysed by crosses with the wild‐type and tetrad dissection. One of the new nuclear suppressor mutants was characterized by temperature‐sensitive growth on non‐fermentable carbon sources. This mutant was transformed with a genomic yeast library. Two independent types of DNA clones were isolated which both complemented the temperature‐sensitive defect. Subcloning and DNA sequencing identified two novel yeast genes which code for proteins with the characteristic features of transcription factors. Both factors exhibit highly structured protein domains consisting of runs and clusters of asparagine and glutamine residues. One of the proteins contains in addition zinc‐finger domains of the C2H2‐type. Therefore the genes are proposed to be named AZF1 ( a sparagine‐rich z inc‐ f finger protein) and PGD1 ( p oly g lutamine d omain protein). Gene disruption of both reading frames has no detectable influence on the vegetative growth on complete glucose or glycerol media, indicating that the genes may act as high copy number suppressors of the mutant defect. Additional transformation experiments showed that AZF1 is also an efficient suppressor for the original defect in the core subunit of mitochondrial RNA polymerase. The DNA sequences for the AZF1 and PGD1 genes were submitted to the EMBL data base (Accession Numbers: Z26253 and Z26254).