WD dipeptide motifs and LXXLL motif of chicken HIRA are essential for interactions with the p48 subunit of chromatin assembly factor-1 and histone deacetylase-2 in vitro and in vivo
WD dipeptide motifs and LXXLL motif of chicken HIRA are essential for interactions with the p48 subunit of chromatin assembly factor-1 and histone deacetylase-2 in vitro and in vivo
复制标题
DOI：
10.1016/j.gene.2004.07.031
复制
发表时间：
2004-11-10
期刊：
影响因子：
3.5
通讯作者：
Nakayama, T
中科院分区：
文献类型：
--
作者：
Ahmad, A;Takami, Y;Nakayama, T
我们克隆了编码鸡HIRA的cDNA，HIRA是酿酒酵母转录辅抑制子Hir 1 p和Hir 2 p的同源物，在其N-末端和C-末端区域分别具有7个WD二肽基序和一个LXXLL基序。它与CAF-1 p48、HDAC-1和2结合，但不与CAF-1 p60、p46多肽和HDAC-3结合。涉及HIRA和CAF-1 p48的截短突变体和错义突变体的免疫沉淀实验显示，不仅HIRA的N-末端半部分中的7个WD二肽基序中的一个对于与CAF-1 p48的相互作用是必需的，而且CAF-1 p48的WD二肽基序对于与HIRA的相互作用也是必需的。这些发现表明HIRA和CAF-1 p48的适当螺旋桨结构对于它们的体外相互作用是必要的。HIRA和HDAC-2的截短突变体和错义突变体的免疫沉淀实验表明，HIRA C端的LXXLL基序和HDAC-2的C端区域是它们在体外相互作用所必需的。此外，HIRA的WD二肽基序和LXXLL基序对于与CAF-1 p48和HDAC-2在体内的相互作用是必需的。两者合计，这些结果表明，HIRA应参与差异，在一些DNA利用过程中，通过其不同的区域与这些蛋白质的相互作用。(C)2004 Elsevier B. V.保留所有权利。
We cloned cDNA encoding chicken HIRA, a homolog of Saccharomyces cerevisiae transcriptional corepressors Hir1p and Hir2p, possessing seven WD dipeptide motifs and a LXXLL motif in its N-terminal and C-terminal regions, respectively. It binds to CAF-1p48, HDAC-I and 2, but not to CAF-1p60, p46 polypeptide and HDAC-3. The immunoprecipitation experiment involving truncated and missense mutants of HIRA and CAF-1p48 revealed not only that even one of seven WD dipeptide motifs in the N-terminal half of HIRA are necessary for the interaction with CAF-1p48, but also that those of CAF-1p48 are necessary for the interaction with HIRA. These findings indicate that the proper propeller structures of both HIRA and CAF-1p48 are necessary for their in vitro interaction. The immumoprecipitation experiment involving truncated and missense mutants of HIRA and HDAC-2 revealed that the LXXLL motif in the C-terminal half of HIRA and a C-terminal region of HDAC-2 are necessary for their in vitro interaction. Moreover, the WD dipeptide motifs and LXXLL motif of HIRA are essential for the interaction with CAF-1p48 and HDAC-2 in vivo. Taken together, these results indicate that HIRA should participate differentially in a number of DNA-utilizing processes through interactions of its distinct regions with these proteins. (C) 2004 Elsevier B.V. All rights reserved.