The ZBTB24-CDCA7 axis regulates HELLS enrichment at centromeric satellite repeats to facilitate DNA methylation

摘要

ZBTB24 regulates methylation of minor satellite DNA in a CDCA7-dependent manner. (A) Southern blot showing hypomethylation of minor satellite DNA in mESCs deficient for or , with deficiency exhibiting more severe effect. (B) Dot blot showing no obvious change in total 5mC level in or mESCs. 3a/3b DKO, / double KO mESCs. (C) Western blot showing the expression of HA-CDCA7 in stable clones generated in (Z4) mESCs. (D) Southern blot analysis of the samples in (C) showing that expression of HA-CDCA7 in mESCs results in recovery of DNA methylation at the minor satellite repeats. (E) Southern blot showing hypomethylation of minor satellite DNA in CH12F3 cells. (F) Dot blot showing comparable 5mC levels in WT and CH12F3 cells. (G) Western blot showing expression of Flag-CDCA7 in CH12F3 cells. Both endogenous (Endo) and Flag-tagged CDCA7 are indicated. (H) Southern blot showing that expression of Flag-CDCA7 in CH12F3 cells rescued DNA methylation at the minor satellite repeats. Relative protein levels in (C and G) were quantified by densitometry using ImageJ, normalized against β-actin