Identification of the binding domains and key amino acids for the interaction of the transcription factors BmPOUM2 and BmAbd-A in Bombyx mori
Identification of the binding domains and key amino acids for the interaction of the transcription factors BmPOUM2 and BmAbd-A in Bombyx mori
复制标题
鉴定家蚕转录因子 BmPOUM2 和 BmAbd-A 相互作用的结合域和关键氨基酸。
DOI：
10.1016/j.ibmb.2016.12.007
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发表时间：
2017-02-01
影响因子：
3.8
通讯作者：
Feng, Qili
中科院分区：
文献类型：
--
作者：
He, Yong;Deng, Huimin;Feng, Qili
转录因子BmPOUM 2与另一个转录因子BmAbd-A相互作用，调控家蚕翅表皮蛋白基因BmWCP 4的表达。本研究报道了BmPOUM 2与BmAbd-A相互作用的结合结构域和氨基酸。鉴定了BmPOUM 2的两种同种型。与全长同种型（BmPOUM 2）相比，短同种型（BmPOUM 2-S）缺少含有POU同源结构域和核定位信号肽（NLS）的114个氨基酸序列。BmPOUM 2和BmPOUM 2-S蛋白均通过POU特异性结构域与BmAbd-A结合。当POU家族基因中高度保守的6个氨基酸（Lys 166、Gly 173、Gln 176、Ser 192、Glu 200和Asn 208）发生突变时，BmPOUM 2不与BmAbd-A结合。BmAbd-A通过同源框结构域或LCR 2（低复杂性区域）结构域与BmPOUM 2相互作用。当同源框和LCR 2结构域中共有的7个氨基酸（Phe 156/248、His 158/250、Ala 175/263、Cys 180/265、Glu 190/268、Trp 196/274和Va 1214/289）突变时，BmAbd-A不与BmPOUM 2结合。BmPOUM 2或BmAbd-A或两者的过表达增加了CHO细胞中BmWCP 4启动子的活性。ChIP和EMSA分析表明，BmAbd-A蛋白与BmWCP 4启动子中的Hox顺式调控元件结合，而BmPOUM 2蛋白与附近的POU CRE结合。提出了BmPOUM 2和BmAbd-A在调控BmWCP 4表达中的相互作用和作用的模型。(C)2016爱思唯尔有限公司版权所有
The transcription factor BmPOUM2 interacted with another transcription factor BmAbd-A to regulate the expression of the wing cuticle protein gene BmWCP4 in Bombyx mori. In this study, the binding domains and amino acids for the interaction between BmPOUM2 and BmAbd-A were reported. Two isoforms of BmPOUM2 were identified. The short isoform (BmPOUM2-S) lacks a 114-amino acid sequence containing a POU-homeodomain and a nuclear localization signal peptide (NLS), as compared to the full-length isoform (BmPOUM2). Both BmPOUM2 and BmPOUM2-S proteins bound to the BmAbd-A through the POU-specific domain. When the six amino acids (Lys166, Gly173, Gln176, Ser192, Glu200 and Asn208) that are highly conserved in POU family genes were mutated, BmPOUM2 did not bind to BmAbd-A. BmAbd-A interacted with BmPOUM2 by the homeobox domain or the LCR2 (low complexity region) domain. When seven amino acids (Phe156/248, His158/250, Ala175/263, Cys180/265, Glu190/268, Trp196/274 and Va1214/289) that are shared in the homeobox and LCR2 domains were mutated, BmAbd-A did not bind to BmPOUM2. Overexpression of either BmPOUM2 or BmAbd-A or both increased the activity of BmWCP4 promoter in CHO cells. ChIP assay and EMSA showed that BmAbd-A protein bound to the Hox cis-regulatory element in the BmWCP4 promoter, while the BmPOUM2 bound to the nearby POU CRE. A model for the interaction and action of BmPOUM2 and BmAbd-A in regulation of the BmWCP4 expression is proposed. (C) 2016 Elsevier Ltd. All rights reserved.