Characterization of the specific interaction between archaeal FHA domain-containing protein and the promoter of a flagellar-like gene-cluster and its regulation by phosphorylation
Characterization of the specific interaction between archaeal FHA domain-containing protein and the promoter of a flagellar-like gene-cluster and its regulation by phosphorylation
复制标题
DOI：
10.1016/j.bbrc.2011.03.011
复制
发表时间：
2011-04-01
影响因子：
3.1
通讯作者：
He, Zheng-Guo
中科院分区：
文献类型：
--
作者：
Duan, Xin;He, Zheng-Guo
含有叉头结构域的转录因子调控的机制和靶基因在古细菌中尚未见报道。本研究利用细菌单杂交技术，首次成功筛选并鉴定了嗜热古菌tokodaisulfolobus tokodaii中含有FHA结构域的潜在转录因子ST0829调控的靶基因。通过体外电泳迁移率转移实验和表面等离子体共振实验，以及体内染色质免疫沉淀实验，我们发现ST0829可以特异性结合古菌鞭毛蛋白编码操纵子ST2519p的启动子区域（包括FlaG、FlaF、FlaH、Flal和flj）。此外，发现含FHA结构域蛋白的磷酸化对其特异性dna结合活性具有负向调节作用。野生型丝氨酸/苏氨酸蛋白激酶ST1565可以抑制ST0829与ST2519p的相互作用，但缺乏激酶活性的突变体ST1565- k166a对ST0829与ST2519p的相互作用没有显著影响。这些发现不仅增加了我们对古菌FHA结构域调控因子功能的认识，而且为进一步了解古菌环境适应的信号机制提供了重要的见解。(C) 2011爱思唯尔公司版权所有。
The mechanism and target genes of regulation by Forkhead (FHA) domain-containing transcription factors have not yet been documented in Archaea. In this study, using a bacterial one-hybrid technique, we successfully screened and identified for the first time a target gene regulated by ST0829, an FHA domain-containing potential transcriptional factor in the hyperthermophilic archaeon Sulfolobus tokodaii. We show that ST0829 could specifically bind to the promoter region of ST2519p, the archaeal flagellar protein-encoding operon (including FlaG, FlaF, FlaH, Flal, and FlaJ) by using both in vitro electrophoretic mobility shift assay and surface plasmon resonance experiments, and in vivo chromatin immunoprecipitation assays. Furthermore, phosphorylation of the FHA domain-containing protein was found to negatively regulate its specific DNA-binding activity. The interaction between ST0829 and ST2519p could be inhibited by wild-type Ser/Thr protein kinase ST1565, but was not significantly affected by its mutant variant ST1565-K166A that lacks kinase activity. These findings not only increase our knowledge about the function of an archaeal FHA domain-containing regulator but also offer important insights for further understanding the signaling mechanism of environmental adaptation in archaea. (C) 2011 Elsevier Inc. All rights reserved.