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Characterization of pUL5, an HCMV protein interacting with the cellular protein IQGAP1.
Virology ( IF 2.8 ) Pub Date : 2019-11-03 , DOI: 10.1016/j.virol.2019.10.018
Giulia Anselmi 1 , Maria Giuliani 1 , Giacomo Vezzani 1 , Rossella Ferranti 1 , Michela Gentile 1 , Mirko Cortese 1 , Diego Amendola 1 , Nicola Pacchiani 1 , Romina D'Aurizio 1 , Luca Bruno 1 , Yasushi Uematsu 1 , Marcello Merola 2 , Domenico Maione 1
Affiliation  

Among the Herpesviridae, human cytomegalovirus (HCMV) owns the largest genome and displays a huge coding potential. Here, we characterized the UL5 gene product (pUL5) of the clinical isolate TR strain. The protein was predicted as a 166-amino-acid membrane protein with a theoretical mass of 19 kDa. Recombinant virus expressing pUL5 with a tag allowed the identification of two pUL5 non-glycosylated species of approximately 19 and 9 kDa, expressed with early and late kinetic respectively. Experiments in infection confirmed that the lower molecular weight species was translated from an internal ATG in the UL5 open reading frame. Confocal microscopy analysis showed that pUL5 localized within the assembly compartment, but is not incorporated in the virion, as shown by Western blot on purified viral particles. Finally, pull-down experiments coupled with mass spectrometry analysis identified IQGAP1 as a pUL5 interactor, giving new hints on possible roles of pUL5 during HCMV infection.

中文翻译:

pUL5的特征,pUL5是与细胞蛋白IQGAP1相互作用的HCMV蛋白。

在疱疹病毒科中,人类巨细胞病毒(HCMV)拥有最大的基因组并显示出巨大的编码潜力。在这里,我们表征了临床分离株TR菌株的UL5基因产物(pUL5)。预测该蛋白质为166个氨基酸的膜蛋白,理论质量为19 kDa。表达带有标签的pUL5的重组病毒可以鉴定出约19和9 kDa的两个pUL5非糖基化物种,分别以早期和晚期动力学表达。感染实验证实,较低分子量的物种是从UL5开放阅读框中的内部ATG转化而来的。共聚焦显微镜分析表明,pUL5定位在装配室内,但未掺入病毒体,如纯化的病毒颗粒上的蛋白质印迹所示。最后,
更新日期:2019-11-04
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