Figure 5: Critical DNA-binding tyrosines are necessary for MuvB recruitment to promoters.
From: Structural basis for LIN54 recognition of CHR elements in cell cycle-regulated promoters
(a) ChIP assay of V5-LIN54 and the indicated mutants after transfection into T98G cells. Following cross-linking and immunoprecipitation with an anti-V5 antibody, quantitative PCR was performed with primers specific to the p107 and CCNB1 promoters. The average promoter enrichment relative to wild-type LIN54 is reported and the error bars are s.d. from three biological replicates and one technical replicate. The P-value evaluating statistical significance was calculated for the promoter enrichment of a mutant relative to wild-type using a two-tailed Student’s t-test. (b) Wild-type or mutant V5-LIN54 were co-transfected into T98G cells with HA-tagged p130, extracts were immunoprecipitated with an anti-V5 antibody and precipitates were probed for the indicated proteins. An IgG band in the LIN9 blot is marked with an asterisk. Triangle corresponds to BMYB band in the input samples. All bands shown in the input for Lin54-V5 are nonspecific as the protein was undetectable until after immunoprecipitation. The full blots are shown in Supplementary Fig. 9.
