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Phosphorylation of tegument protein pp28 contributes to trafficking to the assembly compartment in human cytomegalovirus infection.
Journal of Microbiology ( IF 3 ) Pub Date : 2020-06-27 , DOI: 10.1007/s12275-020-0263-5
Jun-Young Seo 1 , Jin Ah Heo 1 , William J Britt 2
Affiliation  

Human cytomegalovirus (HCMV) UL99 encodes a late tegument protein pp28 that is essential for envelopment and production of infectious virus. This protein is localized to the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) in transfected cells but it localizes to the cytoplasmic assembly compartment (AC) in HCMV-infected cells. Trafficking of pp28 to the AC is required for the assembly of infectious virus. The N-terminal domain (aa 1–61) of pp28 is sufficient for trafficking and function of the wild type protein during viral infection. However, residues required for authentic pp28 trafficking with the exception of the acidic cluster in the N-terminal domain of pp28 remain undefined. Monitoring protein migration on SDS-PAGE, we found that pp28 is phosphorylated in the virus-infected cells and dephosphorylated in the viral particles. By generating substitution mutants of pp28, we showed that three serine residues (aa 41–43) and a tyrosine residue (aa 34) account for its phosphorylation. The mutant forms of pp28 were localized to the plasma membrane as well as the ERGIC in transfected cells. Likewise, these mutant proteins were localized to the plasma membrane as well as the AC in virus-infected cells. These results suggested that phosphorylation of pp28 contributes to its intracellular trafficking and efficient viral assembly and incorporation.

中文翻译:

皮膜蛋白pp28的磷酸化有助于在人类巨细胞病毒感染中转运到装配区室。

人类巨细胞病毒(HCMV)UL99编码后皮蛋白pp28,这对于包膜和生产感染性病毒至关重要。该蛋白位于转染细胞中的内质网-高尔基体中间区室(ERGIC),但位于HCMV感染的细胞中位于细胞质装配区室(AC)。组装感染性病毒需要将pp28贩运到AC。pp28的N末端结构域(aa 1–61)足以在病毒感染期间实现野生型蛋白的运输和功能。但是,除了pp28的N末端域中的酸性簇以外,可靠的pp28转运所需的残基仍然不确定。在SDS-PAGE上监测蛋白质迁移,我们发现pp28在病毒感染的细胞中被磷酸化,而在病毒颗粒中被去磷酸化。通过生成pp28的取代突变体,我们表明三个丝氨酸残基(aa 41-43)和酪氨酸残基(aa 34)导致了其磷酸化。pp28的突变形式位于转染细胞的质膜和ERGIC中。同样,这些突变蛋白也被病毒感染的细胞定位于质膜以及AC。这些结果表明,pp28的磷酸化有助于其细胞内运输以及有效的病毒装配和掺入。这些突变蛋白位于被病毒感染的细胞的质膜以及AC中。这些结果表明,pp28的磷酸化有助于其细胞内运输以及有效的病毒装配和掺入。这些突变蛋白位于被病毒感染的细胞的质膜以及AC中。这些结果表明,pp28的磷酸化有助于其细胞内运输以及有效的病毒装配和掺入。
更新日期:2020-06-27
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