Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

The contribution of pUL74 to growth of human cytomegalovirus is masked in the presence of RL13 and UL128 expression

Laib Sampaio, Kerstin, Stegmann, Cora, Brizic, Ilija, Adler, Barbara, Stanton, Richard James and Sinzger, Christian 2016. The contribution of pUL74 to growth of human cytomegalovirus is masked in the presence of RL13 and UL128 expression. Journal of General Virology 97 , pp. 1917-1927. 10.1099/jgv.0.000475

[img]
Preview
PDF - Published Version
Available under License Creative Commons Attribution.

Download (2MB) | Preview

Abstract

The glycoproteins gH and gL of human cytomegalovirus (HCMV) form a complex either with pUL74 (trimeric complex) or with proteins of the UL128 locus (pentameric complex). While the pentameric complex is dispensable for viral growth in fibroblasts, deletion of pUL74 causes a small plaque phenotype in HCMV lab strains, accompanied by greatly reduced cell-free infectivity. As HCMV isolates shortly after cultivation from clinical specimens do not release cell-free infectious virus, we wondered whether deletion of pUL74 would also affect virus growth in this background. To address this question, we took advantage of the BAC-cloned virus Merlin-RL13tetO that resembles such clinical isolates by growing cell-associated due to inducible expression of the viral RL13 gene. Stop codons were introduced by seamless mutagenesis into UL74 and/or the UL128 locus to abolish expression of the trimeric or pentameric complex, respectively. Virus mutants were reconstituted by transfection of the respective genomes into cultured cells and analyzed regarding focal growth. When the UL128 locus was intact, deletion of pUL74 did not notably affect focal growth of Merlin, irrespective of RL13 expression. In the absence of UL128 expression, foci were increased compared to wild type, and infectious cell-free virus was produced. Under these conditions, disruption of UL74 completely prevented virus spread from initially transfected cells to surrounding cells. In conclusion the contribution of pUL74 is masked when the UL128 locus is expressed at high levels, and its role in cell-free virus spread is only revealed when expression of the pentameric complex is inhibited.

Item Type: Article
Date Type: Published Online
Status: Published
Schools: Medicine
Systems Immunity Research Institute (SIURI)
Subjects: Q Science > QR Microbiology > QR355 Virology
Publisher: Society for General Microbiology
ISSN: 0022-1317
Funders: MRC
Date of First Compliant Deposit: 11 May 2016
Date of Acceptance: 1 April 2016
Last Modified: 02 May 2019 12:28
URI: http://orca-mwe.cf.ac.uk/id/eprint/89017

Citation Data

Cited 7 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item

Downloads

Downloads per month over past year

View more statistics