Li Jian-Feng, Liu Long-Ding, Ma Shao-Hui, Che Yan-Chun, Wang Li-Chun, Dong Cheng-Hong, Zhao Hong-Ling, Liao Yun, Li Qi-Han
Institute of Medical Biology, Chinese Academy of Medical Sciences, Peking Union Medical College, 379 Jiaoling Rd., Kunming, 650118, PR China.
J Biochem. 2004 Aug;136(2):169-76. doi: 10.1093/jb/mvh108.
The interaction between virus and receptor is a process that mimics physiological ligand binding receptors and induces signal transduction. In the investigation of the interaction between HSV1 (Herpes Simplex virus 1) and human fibroblasts via virus binding to its receptor complex on cellular membranes, the HTRP (human transcription regulator protein), a protein encoded by an immediate-early gene of cellular response against the specific stimulation of HSV1 binding, was cloned from a cDNA library established from early gene response mRNA. The localization of HTRP expressed as a fusion polypeptide with a fluorescent protein in HeLa cells was confirmed to be the nucleus. The results of a yeast two-hybrid experiment indicated that HTRP is indeed involved in the interaction with the SAP (mSin3-associate polypeptide) complex via SAP30. A pull-down test and Western blotting in vitro, and immunoprecipitation in vivo also provided evidence in support of this result. The interaction of HTRP with SAP30 in its conserved domain implies that this protein family, as the products of immediate-early genes, comprise functional molecules involved in the transcriptional regulation of cells, which might be related to the inhibition of some cell survival genes.
病毒与受体之间的相互作用是一个模拟生理配体与受体结合并诱导信号转导的过程。在研究单纯疱疹病毒1型(HSV1)与人类成纤维细胞通过病毒与细胞膜上的受体复合物结合的相互作用时,从针对HSV1结合的特异性刺激的细胞早期基因反应mRNA建立的cDNA文库中克隆了HTRP(人类转录调节蛋白),它是由细胞反应的立即早期基因编码的一种蛋白质。在HeLa细胞中,与荧光蛋白融合表达的HTRP的定位被证实是在细胞核中。酵母双杂交实验结果表明,HTRP确实通过SAP30参与了与SAP(mSin3相关多肽)复合物的相互作用。体外下拉试验和蛋白质印迹以及体内免疫沉淀也为这一结果提供了证据。HTRP与SAP30在其保守结构域的相互作用表明,作为立即早期基因的产物,这个蛋白质家族包含参与细胞转录调控的功能分子,这可能与某些细胞存活基因的抑制有关。