The C-terminal domain is the primary determinant of histone H1 binding to chromatin in vivo

被引:187
作者
Hendzel, MJ
Lever, MA
Crawford, E
Th'ng, JPH
机构
[1] Univ Alberta, Cross Canc Inst, Edmonton, AB T6G 1Z2, Canada
[2] Univ Alberta, Dept Oncol, Edmonton, AB T6G 1Z2, Canada
[3] NW Ontario Reg Canc Ctr, Thunder Bay, ON P7B 6V4, Canada
关键词
D O I
10.1074/jbc.M400070200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have used a combination of kinetic measurements and targeted mutations to show that the C-terminal domain is required for high-affinity binding of histone H1 to chromatin, and phosphorylations can disrupt binding by affecting the secondary structure of the C terminus. By measuring the fluorescence recovery after photo-bleaching profiles of green fluorescent protein-histone H1 proteins in living cells, we find that the deletion of the N terminus only modestly reduces binding affinity. Deletion of the C terminus, however, almost completely eliminates histone H1.1 binding. Specific mutations of the C-terminal domain identified Thr-152 and Ser-183 as novel regulatory switches that control the binding of histone H1.1 in vivo. It is remarkable that the single amino acid substitution of Thr-152 with glutamic acid was almost as effective as the truncation of the C terminus to amino acid 151 in destabilizing histone H1.1 binding in vivo. We found that modifications to the C terminus can affect histone H1 binding dramatically but have little or no influence on the charge distribution or the overall net charge of this domain. A comparison of individual point mutations and deletion mutants, when reviewed collectively, cannot be reconciled with simple charge-dependent mechanisms of C-terminal domain function of linker histones.
引用
收藏
页码:20028 / 20034
页数:7
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