Journal of Microbiology

Journal Article

Phosphorylation of tegument protein pp28 contributes to trafficking to the assembly compartment in human cytomegalovirus infection: Jun-Young Seo , Jin Ah Heo , William J. Britt; J. Microbiol. 2020;58(7):624-631. Published online June 27, 2020; DOI: https://doi.org/10.1007/s12275-020-0263-5

Abstract: 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.

Research Support, Non-U.S. Gov't

Removal of Contaminating TEM-la β-Lactamase Gene from Removal of Contaminating TEM-la β-Lactamase Gene from: Jae Seok Song , Jung Hun Lee , Jung-Hyun Lee , Byeong Chul Jeong , Won-Keun Lee , Sang Hee Lee; J. Microbiol. 2006;44(1):126-128.; DOI: https://doi.org/2326 [pii]

Abstract: This study confirms that Taq DNA polymerase could be contaminated with the blaTEM-1a gene. It also proposes two different methods that could be used to overcome DNA contamination: (i) DNase I treatment prior to PCR amplification; and (ii) the use of a highly purified Taq DNA polymerase which was devoid of detectable contamination.

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