Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E

Sinzger, Christian; Hahn, Gabriele; Digel, Margarete; Katona, Ruth; Sampaio, Kerstin Laib; Messerle, Martin; Hengel, Hartmut; Koszinowski, Ulrich H.; Brune, Wolfram; Adler, Barbara

doi:10.1099/vir.0.83286-0

Cited by 365 publications

(364 citation statements)

References 38 publications

Supporting

Mentioning

362

Contrasting

Order By: Relevance

“…The underlying biologic explanations of these patterns could be varied. Cellular tropism is a possible candidate, particularly because the contribution of HCMV glycoproteins to cellular tropism is well-established (80)(81)(82)(83), and genetic variation in genes encoding glycoproteins may play a role in altering HCMV tropism in vivo (27,28). Thus, it is possible that at least a portion of the plasma-enriched SNPs identified in the current study alter HCMV tropism.…”

Section: Discussionmentioning

confidence: 98%

Limits and patterns of cytomegalovirus genomic diversity in humans

Renzette

Pokalyuk

Gibson

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

113

129

View full text Add to dashboard Cite

Human cytomegalovirus (HCMV) exhibits surprisingly high genomic diversity during natural infection although little is known about the limits or patterns of HCMV diversity among humans. To address this deficiency, we analyzed genomic diversity among congenitally infected infants. We show that there is an upper limit to HCMV genomic diversity in these patient samples, with ∼25% of the genome being devoid of polymorphisms. These low diversity regions were distributed across 26 loci that were preferentially located in DNA-processing genes. Furthermore, by developing, to our knowledge, the first genome-wide mutation and recombination rate maps for HCMV, we show that genomic diversity is positively correlated with these two rates. In contrast, median levels of viral genomic diversity did not vary between putatively single or mixed strain infections. We also provide evidence that HCMV populations isolated from vascular compartments of hosts from different continents are genetically similar and that polymorphisms in glycoproteins and regulatory proteins are enriched in these viral populations. This analysis provides the most highly detailed map of HCMV genomic diversity in human hosts to date and informs our understanding of the distribution of HCMV genomic diversity within human hosts.human cytomegalovirus | HCMV | congenital CMV | virology | evolution

show abstract

Section: Discussionmentioning

confidence: 98%

Limits and patterns of cytomegalovirus genomic diversity in humans

Renzette

Pokalyuk

Gibson

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

113

129

View full text Add to dashboard Cite

show abstract

“…The clinical, BAC-derived HCMV isolate, TB40/E-BAC (clone #4; ref (12). ), expressing the SV40-mCherry cassette (13) was utilized for HCMV infections.…”

Section: Methodsmentioning

confidence: 99%

Quantitative Proteomic Discovery of Dynamic Epigenome Changes that Control Human Cytomegalovirus (HCMV) Infection

O’Connor

DiMaggio

Shenk

et al. 2014

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

This work represents the first comprehensive quantitative analysis of global histone post-translational modifications (PTMs) from a virus infection, namely human cytomegalovirus (HCMV) infection. We used a nanoLC-MS/MS platform to identify and quantify the dynamic histone H3 and H4 PTMs expressed during HCMV replication in primary fibroblasts. Specifically, we examined the changes in histone PTMs over a 96 h time course to sample the immediate early (IE), early (E), and late (L) stages of viral infection. Several changes in histone H3 and H4 PTMs were observed, including a marked increase in H3K79me2 and H3K27me3K36me2, and a decrease in H4K16ac, highlighting likely epigenetic strategies of transcriptional activation and silencing during HCMV lytic infection. Heavy methyl-SILAC (hm-SILAC) was used to further confirm the histone methylation flux (especially for H3K79) during HCMV infection. We evaluated DOT1L (the H3K79 methyltransferase) mRNA levels in mock and HCMV-infected cells over a 96 h time course, and observed a significant increase in this methyltransferase as early as 24 hpi showing that viral infection up-regulates DOT1L expression, which drives H3K79me2. We then used shRNA to create a DOT1L knockdown cell population, and found that HCMV infection of the knockdown cells resulted in a 10-fold growth defect when compared with infected control cells not subjected to knockdown. This work documents multiple histone PTMs that occur in response to HCMV infection of fibroblasts, and provides a framework for evaluation of the role of epigenetic modifications in the virus-host interaction. Molecular & Cellular Proteomics

show abstract

“…While TB40/E encodes functional versions of the antiapoptotic proteins vMIA, viral inhibitor of caspase activation (vICA) (Skaletskaya et al, 2001) and pUL38 (Moorman et al, 2008;Terhune et al, 2007) and carries additional genes in the UL-b9 genomic region (Sinzger et al, 2008), AD encodes a nonfunctional vICA (Skaletskaya et al, 2001) and contains a longer UL-b9 region. Inclusion of all three strains thus allowed us to evaluate the contribution of the US2-US11 genes in protection from cell death in a vMIA+/pUL38+/vICA2 genetic background (AD) and to compare the behaviour of AD to that of TB40/E, a virus carrying a more complete set of genes (Sinzger et al, 2008).…”

Section: Presence Of the Us2-us11 Genomic Region Confers Protection Fmentioning

confidence: 99%

Human cytomegalovirus US9 protein contains an N-terminal signal sequence and a C-terminal mitochondrial localization domain, and does not alter cellular sensitivity to apoptosis

Mandic

Miller

Coulter

et al. 2009

Journal of General Virology

View full text Add to dashboard Cite

The human cytomegalovirus (CMV) US2-US11 genomic region contains a cluster of genes whose products interfere with antigen presentation by the major histocompatibility complex (MHC) proteins. Although included in this cluster, the US9 gene encodes a glycoprotein that does not affect MHC activity and whose function is still largely uncharacterized. An in silico analysis of the US9 amino-acid sequence uncovered the presence of an N-terminal signal sequence (SS) and a C-terminal transmembrane domain containing the specific hallmarks of known mitochondrial localization sequences (MLS). Expression of full-length US9 and of US9 deletion mutants fused to GFP revealed that the N-terminal SS mediates US9 targeting to the endoplasmic reticulum (ER) and that the C-terminal MLS is both necessary and sufficient to direct US9 to mitochondria in the absence of a functional SS. This dual localization suggested a possible role for US9 in protection from apoptosis triggered by ER-to-mitochondria signalling. Fibroblasts infected with the US2-US11 deletion mutant virus RV798 or with the parental strain AD169varATCC were equally susceptible to death triggered by exposure to tumour necrosis factor (TNF)-a, tunicamycin, thapsigargin, brefeldin A, lonidamine and carbonyl cyanide m-chloro phenyl hydrazone, but were 1.6-fold more sensitive to apoptosis induced by hygromycin B. Expression of US9 in human embryonic kidney 293T cells or in fibroblasts, however, did not protect cells from hygromycin B-mediated death. Together, these results classify US9 as the first CMV-encoded protein to contain an N-terminal SS and a C-terminal MLS, and suggest a completely novel role for this protein during infection. INTRODUCTIONHuman cytomegalovirus (CMV) is a very prevalent betaherpesvirus that poses serious medical concerns in immunocompromised individuals . The ability to evade or modulate host immune responses, coupled with the capacity to establish latency after primary infection, is at the basis of the tremendous success of CMV as a human pathogen.The US2-US11 genomic region has been shown to be dispensable for viral replication in primary human fibroblasts (HF) (Jones & Muzithras, 1992;Kollert-Jons et al., 1991), and to encode five endoplasmic reticulum (ER)-resident glycoproteins, US2, US3, US6, US10 and US11, that disrupt antigen presentation by the major histocompatibility complex (MHC) class I and class II proteins (Lin et al., 2007). Despite sharing some very limited homology with US6, US10 and US11, US9 does not bind to or reduce MHC cell surface levels (Hegde et al., 2002(Hegde et al., , 2006Pereira et al., 1995). US9 was originally described as a member of the US6 family of proteins, which includes the products of the US6-US11 genes. These proteins contain markedly hydrophobic sequences at both the N-and C-termini, and were predicted to insert into cellular membranes (Weston & Barrell, 1986). US9 intracellular localization was initially examined in nonpolarized Madin-Darby canine kidney cells constitutively expressing a C-terminal-tagged ...

show abstract

Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E

Cited by 365 publications

References 38 publications

Limits and patterns of cytomegalovirus genomic diversity in humans

Limits and patterns of cytomegalovirus genomic diversity in humans

Quantitative Proteomic Discovery of Dynamic Epigenome Changes that Control Human Cytomegalovirus (HCMV) Infection

Human cytomegalovirus US9 protein contains an N-terminal signal sequence and a C-terminal mitochondrial localization domain, and does not alter cellular sensitivity to apoptosis

Contact Info

Product

Resources

About