FIG. 3.

Localization of the mutations in rsp5-25. (A) The rsp5-25 mutation was mapped by using a plasmid that contains the wild-type RSP5 allele, digested with indicated restriction enzymes for use in plasmid gap repair experiments. The gapped regions are indicated as thick lines. After transformation into an rsp5-25 strain, plasmid gap repair results in the copying of the corresponding genomic region onto the plasmid. The results of complementation experiments in rad1 rad52 rfa1-D228Y rsp5-25 are depicted. The black boxes on the RSP5 ORF depict the WW domains, and the hatched box represents the ubiquitination domain of the protein. (B) Localization of the two rsp5-25 mutations in the ubiquitination domain. The mutated tryptophan residue is indicated as a boldface letter, and the ochre codon is indicated by an asterisk. The protein sequences of several Rsp5 homologs are also shown to indicate the conservation near the mutated residues.