Functional domains of the ubiquitous chromatin protein DEK
Functional domains of the ubiquitous chromatin protein DEK
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DOI：
10.1128/mcb.24.13.6000-6010.2004
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发表时间：
2004-07-01
影响因子：
5.3
通讯作者：
Waldmann, T
中科院分区：
文献类型：
--
作者：
Kappes, F;Scholten, I;Waldmann, T
DEK最初被描述为原癌基因蛋白，现在已知是后生动物染色质的主要成分。DEK能够通过引入超螺旋来改变DNA的结构。为了寻找DEK的相互作用伙伴和功能结构域，我们进行了酵母双杂交筛选和突变分析。双杂交筛选产生了DEK的C-末端片段，表明DEK能够多聚化。我们可以将该结构域定位于氨基酸270至350，并表明多聚化依赖于体外CK 2激酶的磷酸化。我们还发现了DEK的两个DNA结合结构域，一个在包括氨基酸87至187的片段上，并且包含SAF盒DNA结合基序，其位于氨基酸149和187之间。该区域足以将超螺旋引入DNA中。第二DNA结合结构域位于氨基酸270和350之间，因此与多聚化结构域重叠。我们发现，这两个DNA相互作用的结构域不同的结合特性和他们的能力，以响应CK 2磷酸化。
DEK was originally described as a proto-oncogene protein and is now known to be a major component of metazoan chromatin. DEK is able to modify the structure of DNA by introducing supercoils. In order to find interaction partners and functional domains of DEK, we performed yeast two-hybrid screens and mutational analyses. Two-hybrid screening yielded C-terminal fragments of DEK, suggesting that DEK is able to multimerize. We could localize the domain to amino acids 270 to 350 and show that multimerization is dependent on phosphorylation by CK2 kinase in vitro. We also found two DNA binding domains of DEK, one on a fragment including amino acids 87 to 187 and containing the SAF-box DNA binding motif, which is located between amino acids 149 and 187. This region is sufficient to introduce supercoils into DNA. The second DNA binding domain is located between amino acids 270 and 350 and thus overlaps the multimerization domain. We show that the two DNA-interacting domains differ in their binding properties and in their abilities to respond to CK2 phosphorylation.