FIG 5.

Mutations of the FCD domains of FRQ cause temperature-sensitive FRQ-CK1a association and partial loss of temperature compensation. (A to C, left) Representative photos of race tubes used to evaluate conidiation rhythms of the wild-type (frq complementation strain) and FCD mutant strains. (Right) Plot of period versus temperature for wild-type and FCD mutant strains. Error bars are standard errors of means (n = 5 to 9). (D, left) Western blot analyses of FRQ immunoprecipitated with anti-CK1a antibody from extracts of the wild-type and Q494N strains grown in constant light at 21 and 29°C. (Right) Plot of relative amount of FRQ in wild-type and Q494N strains. Quantification of relative FRQ-CK1a interaction levels is based on the ratio of IP to input and normalized to CK1a level. Error bars are standard deviations (n = 3). ns, not significant; *, P < 0.05; Student’s t test. (E, left) Western blot analyses of FRQ in the wild-type and Q494N strains grown in constant light at 21 or 29°C for the indicated number of hours after CHX addition. (Right) Plot of relative FRQ levels at the indicated hours after CHX treatment. Error bars are standard deviations (n = 3). *, P < 0.05; **, P < 0.01; Student’s t test.