Cloning, sequencing, disruption and phenotypic analysis of uvsC, an Aspergillus nidulans homologue of yeast RAD51.
We have cloned the uvsC gene of Aspergillus nidulans by complementation of the A. nidulans uvsC114 mutant. The predicted protein UVSC shows 67.4% sequence identity to the Saccharomyces cerevisiae Rad51 protein and 27.4% sequence identity to the Escherichia coli RecA protein. Transcription of uvsC is induced by methyl-methane sulphonate (MMS), as is transcription of RAD51 of yeast. Similar levels of uvsC transcription were observed after MMS induction in a uvsC+ strain and the uvsC114 mutant. The coding sequence of the uvsC114 allele has a deletion of 6 bp, which results in deletion of two amino acids and replacement of one amino acid in the translation product. In order to gain more insight into the biological function of the uvsC gene, a uvsC null mutant was constructed, in which the entire uvsC coding sequence was replaced by a selectable marker gene. Meiotic and mitotic phenotypes of a uvsC+ strain, the uvsC114 mutant and the uvsC null mutant were compared. The uvsC null mutant was more sensitive to both UV and MMS than the uvsC114 mutant. The uvsC114 mutant arrested in meiotic prophase-I. The uvsC null mutant arrested at an earlier stage, before the onset of meiosis. One possible interpretation of these meiotic phenotypes is that the A. nidulans homologue of Rad51 of yeast has a role both in the specialized processes preceding meiosis and in meiotic prophase I.
van Heemst
D
D
Department of Genetics, Agricultural University Wageningen, The Netherlands.
Swart
K
K
Holub
E F
EF
van Dijk
R
R
Offenberg
H H
HH
Goosen
T
T
van den Broek
H W
HW
Heyting
C
C
D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D001483:Base Sequence; D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D005656:Fungal Proteins / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D008540:Meiosis; D008741:Methyl Methanesulfonate / Q000633:toxicity; D008938:Mitosis; D008969:Molecular Sequence Data; D009153:Mutagens; D009154:Mutation; D010641:Phenotype; D051135:Rad51 Recombinase; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D014466:Ultraviolet Rays
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D009153:Mutagens; C107165:UvsC protein, Emericella nidulans; D008741:Methyl Methanesulfonate; D051135:Rad51 Recombinase