The yeast HRS1 gene encodes a polyglutamine-rich nuclear protein required for spontaneous and hpr1-induced deletions between direct repeats.
The hrs1-1 mutation was isolated as an extragenic suppressor of the hyperrecombination phenotype of hpr1 delta cells. We have cloned, sequenced and deleted from the genome the HRS1 gene. The DNA sequence of the HRS1 gene reveals that it is identical to PGD1, a gene with no reported function, and that the Hrs1p protein contains polyglutamine stretches typically found in transcription factors. We have purified a His(6) tagged version of Hrs1p protein from E. coli and have obtained specific anti-Hrs1p polyclonal antibodies. We show that Hrs1p is a 49-kD nuclear protein, as determined by indirect immunofluorescence microscopy and Western blot analysis. The hrs1 delta null mutation reduces the frequency of deletions in wild-type and hpr1 delta backgrounds sevenfold below wild-type and rad52 levels. Furthermore, hrs1 delta cells show reduced induction of the GAL1,10 promoter relative to wild-type cells. Our results suggest that Hrs1p is required for the formation of deletions between direct repeats and that it may function in gene expression. This suggests a connection between gene expression and direct repeat recombination. In this context, we discuss the possible roles of Hrs1p and Hpr1p in initiation of direct-repeat recombination.
Santos-Rosa
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H
Departamento de Genetica, Universidad de Sevilla, Spain.
Clever
B
B
Heyer
W D
WD
Aguilera
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A
D000595:Amino Acid Sequence; D000818:Animals; D000908:Antibodies, Fungal / Q000302:isolation & purification; D000915:Antibody Affinity; D001483:Base Sequence; D004260:DNA Repair; D004271:DNA, Fungal; D019084:Fluorescent Antibody Technique, Indirect; D005656:Fungal Proteins / Q000235:genetics* / Q000276:immunology; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D008540:Meiosis; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000276:immunology; D010455:Peptides; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051381:Rats; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics*; D014466:Ultraviolet Rays
D000908:Antibodies, Fungal; D004271:DNA, Fungal; D005656:Fungal Proteins; D009687:Nuclear Proteins; D010455:Peptides; C097188:polyglutamine