FIGURE 7:

Mgm101^Y139A is functionally defective in vivo but competent in ssDNA binding in vitro. (A) Sequence alignment of Mgm101 with human Rad52. Shown in the Mgm101 functional core is Y139, which is equivalent to Y47 in yRad52 (yeast), Y65 in hRad52 (humans), and Y42 in Sak (lactococcal ul36). (B) Meiotic analysis showing that the mgm101^Y139A is unable to complement the mgm101Δ::kan allele for growth on complete medium containing glycerol and ethanol (Gly+EtOH) as carbon sources. Four complete tetrads are shown, in which the segregants cosegregating mgm101Δ::kan (G418^R) and the mgm101^Y139A allele (Ura⁺) are circled. (C) Size exclusion chromatography of purified Mgm101^Y139A on a Superose 6 column showing that the mutant variant coelutes with the wild-type protein. (D) ssDNA-binding activity of Mgm101^Y139A compared with wild type. Error bars, average deviations of three independent experiments.