Histone recycling function of Mrc1 is conserved in mammalian cells
(A) (top) Mouse CLASPIN with annotated domains (BP1 and BP2: basic patch 1 and 2, CKBD: Chk1 binding domain, AP: acidic patch) and alignment with Polα and Mcm2 HBDs. (Bottom) overview of mutation in CLASPINΔYY.
(B) WB analysis of WT and CLASPIN mutant mESCs.
(C) Cell-cycle distribution based on mean EdU intensity and total DAPI intensity of WT and CLASPIN mutant mESCs.
(D) High-content microscopy of mean EdU intensity in WT and CLASPIN mutant mESCs.
(E) Average SCAR-seq profile of H3K27me3 and H4K20me0 in WT and CLASPIN mutant mESCs. (C–E) n = 2 biological replicates.
(F) Model: Mrc1 acts as a central coordinator of histone-based inheritance though its ability to bind and transfer H3-H4 tetramers to both leading and lagging strands, with the latter involving joint histone binding with Mcm2 to facilitate transfer to Polα and the lagging strand.