Characterisation of Schizosaccharomyces pombe rad31, a UBA-related gene required for DNA damage tolerance.
The fission yeast rad31-1 mutant is sensitive to both UV and ionising radiation and exhibits a growth defect at 35 degrees C. In addition, the mutant displays defects in cell morphology and nuclear division at 26 degrees C which are exaggerated at 35 degrees C. We have cloned the rad31 gene and have shown that it is not essential for viability, although cells containing a disrupted rad31 gene grow slowly. The null allele has similar cell and nuclear morphologies to the original allele and displays an extremely high frequency of loss of minichromosomes. rad31 is not required for either the S/M or G2/M checkpoint, however double mutant analysis indicates that rad31 acts in a process which is defective in the checkpoint rad mutants and which involves hus5 . Sequence analysis indicates that rad31 encodes a protein which is related to ubiquitin activating proteins and more particularly to an ORF in Saccharomyces cerevisiae and to the Arabidopsis thaliana AXR1 and human APP-BP1 genes. We have isolated the S.cerevisiae sequence, which we have named RHC31 ( ad31homologue in S. erevisiae), since we show that it can complement the slow growth phenotype and radiation sensitivity of S.pombe rad31.
Shayeghi
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Biochemistry, School of Biological Sciences, University of Sussex, Falmer, Brighton BN1 9QG, UK.
Doe
C L
CL
Tavassoli
M
M
Watts
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FZ
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins; D015825:Chromosomes, Fungal / Q000528:radiation effects; D003001:Cloning, Molecular; D004249:DNA Damage / Q000235:genetics* / Q000528:radiation effects; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000235:genetics; D004307:Dose-Response Relationship, Radiation; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D005720:Gamma Rays; D006133:Growth Substances; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000235:genetics; D016366:Open Reading Frames; D010940:Plant Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects; D029702:Schizosaccharomyces pombe Proteins; D014466:Ultraviolet Rays
C081977:AXR1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D006133:Growth Substances; D009419:Nerve Tissue Proteins; D010940:Plant Proteins; C105272:RAD31 protein, S pombe; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D029702:Schizosaccharomyces pombe Proteins