A fragment of the yeast DNA repair protein Rad4 confers toxicity to E. coli and is required for its interaction with Rad7 protein.
The RAD4 gene of Saccharomyces cerevisiae is required for the incision of damaged DNA during nucleotide excision repair. Plasmids carrying the wild-type RAD4 gene cannot be propagated in Escherichia coli. In this study, a rad4 mutant that can be grown in E. coli was isolated. This rad4 allele is deleted of a large positively charged segment of the RAD4 coding region which is toxic to E. coli when expressed alone. The deletion mutant retains its ability to interact with Rad23 protein but not with Rad7 protein and is defective in nucleotide excision repair. The smallest Rad4 fragment that is toxic to E. coli consists of 336 amino acids with a calculated pI=9.99.
Wei
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Laboratory of Molecular Pathology, Department of Pathology, University of Texas Southwestern Medical Center, Dallas, TX 75235, USA.
Friedberg
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EC
D000483:Alleles; D000595:Amino Acid Sequence; D004260:DNA Repair / Q000235:genetics*; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics* / Q000502:physiology* / Q000633:toxicity; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics* / Q000502:physiology / Q000633:toxicity; D010641:Phenotype; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D011503:Transglutaminases
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D010446:Peptide Fragments; C082463:RAD4 protein, S pombe; C063915:RAD7 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D011503:Transglutaminases