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Genetics, Vol 142, 705-716, Copyright © 1996
INVESTIGATIONS |
The Yeast HRS1 Gene Encodes a Polyglutamine-Rich Nuclear Protein Required for Spontaneous and hpr1-Induced Deletions Between Direct Repeats
H. Santos-Rosa, B. Clever, W. D. Heyer and A. Aguilera
Departamento de Genetica, Facultad de Biologia, Universidad de Sevilla, E-41012 Sevilla, Spain
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.
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