Disruptions of the Ustilago maydis REC2 gene identify a protein domain important in directing recombinational repair of DNA

The REC2 gene of Ustilago maydis encodes a homologue of the Escherichia coli RecA protein and was first identified in a screen for UV‐sensitive mutants. The original isolate, rec2‐1, was found to be deficient in repair of DNA damage, genetic recombination and meiosis. We report here that the rec2‐19...

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Veröffentlicht in:Molecular microbiology 2001-06, Vol.40 (6), p.1415-1426
Hauptverfasser: Kojic, Milorad, Thompson, Chad W., Holloman, William K.
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Thompson, Chad W.
Holloman, William K.
description The REC2 gene of Ustilago maydis encodes a homologue of the Escherichia coli RecA protein and was first identified in a screen for UV‐sensitive mutants. The original isolate, rec2‐1, was found to be deficient in repair of DNA damage, genetic recombination and meiosis. We report here that the rec2‐197 allele, which was constructed by gene disruption, retains some biological activity and is partially dominant with respect to REC2. The basis for the residual activity is probably as a result of expression of a diffusible product from the rec2‐197 allele that augments or interferes with REC2 functions. This product appears to be a polypeptide expressed from a remnant of the 5′ end of the open reading frame that was not removed in creating the gene disruption. The mutator activity and disturbed meiosis of rec2‐197 suggest that the Rec2 protein functions in a process that avoids spontaneous mutation and insures faithful meiotic chromosome segregation. A prediction based on the phenotype of rec2‐197 is that Rec2 protein interacts with one or more other proteins in directing these functions. To identify interacting proteins we performed a yeast two‐hybrid screen and found Rad51 as a candidate. Rec2‐197 and Rad51 appear to interact to a similar degree.
doi_str_mv 10.1046/j.1365-2958.2001.02490.x
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The original isolate, rec2‐1, was found to be deficient in repair of DNA damage, genetic recombination and meiosis. We report here that the rec2‐197 allele, which was constructed by gene disruption, retains some biological activity and is partially dominant with respect to REC2. The basis for the residual activity is probably as a result of expression of a diffusible product from the rec2‐197 allele that augments or interferes with REC2 functions. This product appears to be a polypeptide expressed from a remnant of the 5′ end of the open reading frame that was not removed in creating the gene disruption. The mutator activity and disturbed meiosis of rec2‐197 suggest that the Rec2 protein functions in a process that avoids spontaneous mutation and insures faithful meiotic chromosome segregation. A prediction based on the phenotype of rec2‐197 is that Rec2 protein interacts with one or more other proteins in directing these functions. To identify interacting proteins we performed a yeast two‐hybrid screen and found Rad51 as a candidate. 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The original isolate, rec2‐1, was found to be deficient in repair of DNA damage, genetic recombination and meiosis. We report here that the rec2‐197 allele, which was constructed by gene disruption, retains some biological activity and is partially dominant with respect to REC2. The basis for the residual activity is probably as a result of expression of a diffusible product from the rec2‐197 allele that augments or interferes with REC2 functions. This product appears to be a polypeptide expressed from a remnant of the 5′ end of the open reading frame that was not removed in creating the gene disruption. The mutator activity and disturbed meiosis of rec2‐197 suggest that the Rec2 protein functions in a process that avoids spontaneous mutation and insures faithful meiotic chromosome segregation. A prediction based on the phenotype of rec2‐197 is that Rec2 protein interacts with one or more other proteins in directing these functions. 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subjects Amino Acid Sequence
Binding Sites
Chromosome Segregation
Diploidy
DNA Repair - drug effects
DNA Repair - genetics
DNA Repair - radiation effects
Fungal Proteins - genetics
Fungal Proteins - metabolism
Homozygote
Meiosis
Methyl Methanesulfonate - pharmacology
Molecular Sequence Data
Mutagens - pharmacology
Mutation
Protein Structure, Tertiary
Rad51 protein
REC2 gene
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
Recombination, Genetic
Spores, Fungal - physiology
Two-Hybrid System Techniques
Ultraviolet Rays
Ustilago - drug effects
Ustilago - physiology
Ustilago - radiation effects
Ustilago maydis
Yeasts - genetics
title Disruptions of the Ustilago maydis REC2 gene identify a protein domain important in directing recombinational repair of DNA
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