Cse4 Is Part of an Octameric Nucleosome in Budding Yeast

被引:136
作者
Camahort, Raymond [1 ,2 ]
Shivaraju, Manjunatha [1 ,3 ]
Mattingly, Mark [1 ]
Li, Bing [1 ,4 ]
Nakanishi, Shima [1 ]
Zhu, Dongxiao [1 ,5 ]
Shilatifard, Ali [1 ]
Workman, Jerry L. [1 ]
Gerton, Jennifer L. [1 ,2 ]
机构
[1] Stowers Inst Med Res, Kansas City, MO 64110 USA
[2] Univ Kansas, Med Ctr, Dept Biochem & Mol Biol, Kansas City, KS 66160 USA
[3] Open Univ, Milton Keynes MK7 6BJ, Bucks, England
[4] Univ Texas SW Med Ctr Dallas, Dept Mol Biol, Dallas, TX 75390 USA
[5] Univ New Orleans, Dept Comp Sci, New Orleans, LA 70148 USA
基金
美国国家卫生研究院;
关键词
A CENP-A; HISTONE H3; IN-VIVO; CENTROMERE; DNA; CHROMATIN; VARIANT; SCM3; SEQUENCE; CORE;
D O I
10.1016/j.molcel.2009.07.022
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
The budding yeast CenH3 histone variant Cse4 localizes to centromeric nucleosomes and is required for kinetochore assembly and chromosome segregation. The exact composition of centromeric Cse4-containing nucleosomes is a subject of debate. Using unbiased biochemical, cell-biological, and genetic approaches, we have tested the composition of Cse4-containing nucleosomes. Using micrococcal nuclease-treated chromatin, we find that Cse4 is associated with the histones H2A, H2B, and H4, but not H3 or the nonhistone protein Scm3. Overexpression of Cse4 rescues the lethality of a scm3 deletion, indicating that Scm3 is not essential for the formation of functional centromeric chromatin. We also find that octameric Cse4 nucleosomes can be reconstituted in vitro. Furthermore, Cse4-Cse4 dimerization occurs in vivo at the centromeric nucleosome, and this requires the predicted Cse4-Cse4 dimerization interface. Taken together, our experimental evidence supports the model that the Cse4 nucleosome is an octamer, containing two copies each of Cse4, H2A, H2B, and H4.
引用
收藏
页码:794 / 805
页数:12
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