Dengue Virus Induces Novel Changes in Gene Expression of Human Umbilical Vein Endothelial Cells

Warke, Rajas V.; Xhaja, Kris; Martin, Katherine J.; Fournier, Marcia F.; Shaw, Sunil K.; Brizuela, Nathaly; Bosch, Norma B. de; Lapointe, David S.; Ennis, Francis A.; Rothman, Alan L.; Bosch, Irene

doi:10.1128/jvi.77.21.11822-11832.2003

Cited by 140 publications

(112 citation statements)

References 62 publications

Supporting

Mentioning

101

Contrasting

Unclassified

Order By: Relevance

“…The host cell response to flavivirus infection has been characterized in several studies (10,27,40,45). As might be expected, many of the host genes differentially regulated during infection are proinflammatory genes, such as cytokines or interferon response factors.…”

Section: Discussionmentioning

confidence: 96%

The Src Family Kinase c-Yes Is Required for Maturation of West Nile Virus Particles

Hirsch¹,

Medigeshi²,

Meyers³

et al. 2005

J Virol

View full text Add to dashboard Cite

The role of cellular genes in West Nile virus (WNV) replication is not well understood. Examination of cellular transcripts upregulated during WNV infection revealed an increase in the expression of the src family kinase (SFK) c-Yes. WNV-infected cell lines treated with the SFK inhibitor PP2 demonstrated a 2-to 4-log decrease in viral titers, suggesting that SFK activity is required for completion of the viral replication cycle. RNA interference mediated knock-down of c-Yes, but not c-Src, and similarly reduced virus yield, specifically implicating c-Yes in WNV production. Interestingly, PP2 treatment did not reduce intracellular levels of either viral RNA or protein, suggesting that the drug does not act on the early stages of replication. However, endoglycosidase H (endoH) digestion of the viral envelope (E) glycoprotein revealed that the acquisition of endoH-resistant glycans by E, but not endogenous major histocompatibility complex class I, was reduced in PP2-treated cells, demonstrating that E specifically does not traffic beyond the endoplasmic reticulum in the absence of SFK activity. Electron microscopy further revealed that PP2-treated WNV-infected cells accumulated an increased number of virions in the ER compared to untreated cells. Therefore, we conclude that inhibition of SFK activity did not interfere with virus assembly but prevented transit of virions through the secretory pathway. These results identify c-Yes as a cellular protein that is involved in WNV assembly and egress.West Nile virus (WNV) is a newly emerging pathogen that has become a significant threat to the U.S. population. First detected in Uganda in 1937, the virus appeared in the United States in the summer of 1999, and since that time it has reemerged each year over an increasing geographical area (1). Typical of flaviviruses, WNV contains a single-stranded positive-sense 11-kb RNA genome. The genome is translated into a single polyprotein, which is cleaved by host and virus-encoded proteases into 10 functional subunits (5, 20). The WNV particle is composed of the subunits derived from the amino terminus of the polyprotein: capsid (C), precursor membrane/ membrane (prM/M), and envelope (E). The remaining seven proteins are nonstructural and function in virus replication. WNV infection results in proliferation and reorganization of intracellular membranes into several unique structures that colocalize with specific subsets of viral proteins, suggesting a distinct spatial segregation of the stages of virus replication (24, 48).WNV virion assembly appears to occur at the rough endoplasmic reticulum (ER), presumably by budding of the nucleocapsid (NC) into the ER lumen (5,20,28). In the context of viral infection, packaging of the genomic RNA requires NS2A and the replication of the RNA by the viral RNA-dependent RNA polymerase NS5 (16,21). The mechanism of NC budding into the ER and acquisition of the viral membrane and glycoproteins is unknown, although a hydrophobic region in the C protein has been proposed to be important in these ...

show abstract

Section: Discussionmentioning

confidence: 96%

The Src Family Kinase c-Yes Is Required for Maturation of West Nile Virus Particles

Hirsch¹,

Medigeshi²,

Meyers³

et al. 2005

J Virol

View full text Add to dashboard Cite

show abstract

“…since in these cells there is active DENV infection but there is a clear difference in the level of virus production between WT and SK1 2/2 cells. Global gene expression analysis showed expected DENV-induced responses in WT iMEFs, including induction of chemokines, pathogen recognition pathways and ISGs (Conceição et al, 2010;Fink et al, 2007;Liew & Chow, 2006;Suthar et al, 2013;Warke et al, 2003). Interestingly, DENV infection of WT iMEFs also induced mRNAs for a number of SP100-related, PARPrelated and Schlafen proteins, which may be of future interest.…”

Section: Discussionmentioning

confidence: 99%

Reduction in sphingosine kinase 1 influences the susceptibility to dengue virus infection by altering antiviral responses

Clarke

Davies

Calvert

et al. 2016

Journal of General Virology

View full text Add to dashboard Cite

Sphingosine kinase (SK) 1 is a host kinase that enhances some viral infections. Here we investigated the ability of SK1 to modulate dengue virus (DENV) infection in vitro. Overexpression of SK1 did not alter DENV infection; however, targeting SK1 through chemical inhibition resulted in reduced DENV RNA and infectious virus release. DENV infection of SK1 2/2 murine embryonic fibroblasts (MEFs) resulted in inhibition of infection in an immortalized line (iMEF) but enhanced infection in primary MEFs (18MEFs). Global cellular gene expression profiles showed expected innate immune mRNA changes in DENV-infected WT but no induction of these responses in SK1 2/2 iMEFs. Reverse transciption PCR demonstrated a low-level induction of IFN-b and poor induction of mRNA for the interferon-stimulated genes (ISGs) viperin, IFIT1 and CXCL10 in DENV-infected SK1 2/2 compared with WT iMEFs. Similarly, reduced induction of ISGs was observed in SK1 2/2 18MEFs, even in the face of high-level DENV replication. In both iMEFs and 18MEFs, DENV infection induced production of IFN-b protein. Additionally, higher basal levels of antiviral factors (IRF7, CXCL10 and OAS1) were observed in uninfected SK1 2/2 iMEFs but not 18MEFs. This suggests that, in this single iMEF line, lack of SK1 upregulates the basal levels of factors that may protect cells against DENV infection. More importantly, regardless of the levels of DENV replication, all cells that lacked SK1 produced IFN-b but were refractory to induction of ISGs such as viperin, IFIT1 and CXCL10. Based on these findings, we propose new roles for SK1 in affecting innate responses that regulate susceptibility to DENV infection.

show abstract

“…Cell samples were harvested in duplicate at three time points, 3, 5, and 7 days, after seeding in lrECM. Purified total cellular RNA was biotin labeled and hybridized to human oligonucleotide microarrays (Affymetrix HG-U133A; Affymetrix, Santa Clara, CA) as described previously (20). Experiments with Affymetrix-present P-call rates of >30% were included in the analysis.…”

Section: Methodsmentioning

confidence: 99%

Gene Expression Signature in Organized and Growth-Arrested Mammary Acini Predicts Good Outcome in Breast Cancer

et al. 2006

Self Cite

View full text Add to dashboard Cite

Nonmalignant human mammary epithelial cells (HMEC) seeded in laminin-rich extracellular matrix (lrECM) form polarized acini and, in doing so, transit from a disorganized proliferating state to an organized growth-arrested state. We hypothesized that the gene expression pattern of organized and growth-arrested HMECs would share similarities with breast tumors with good prognoses. Using Affymetrix HG-U133A microarrays, we analyzed the expression of 22,283 gene transcripts in 184 ( finite life span) and HMT3522 S1 (immortal nonmalignant) HMECs on successive days after seeding in a lrECM assay. Both HMECs underwent growth arrest in G 0 -G 1 and differentiated into polarized acini between days 5 and 7. We identified gene expression changes with the same temporal pattern in both lines and examined the expression of these genes in a previously published panel of microarray data for 295 breast cancer samples. We show that genes that are significantly lower in the organized, growth-arrested HMEC than in their proliferating counterparts can be used to classify breast cancer patients into poor and good prognosis groups with high accuracy. This study represents a novel unsupervised approach to identifying breast cancer markers that may be of use clinically. (Cancer Res 2006; 66(14): 7095-102)

show abstract

Dengue Virus Induces Novel Changes in Gene Expression of Human Umbilical Vein Endothelial Cells

Abstract: Endothelial cells are permissive to dengue virus (DV) infection in vitro, although their importance as targets of DV infection in vivo remains

Cited by 140 publications

References 62 publications

The Src Family Kinase c-Yes Is Required for Maturation of West Nile Virus Particles

The Src Family Kinase c-Yes Is Required for Maturation of West Nile Virus Particles

Reduction in sphingosine kinase 1 influences the susceptibility to dengue virus infection by altering antiviral responses

Gene Expression Signature in Organized and Growth-Arrested Mammary Acini Predicts Good Outcome in Breast Cancer

Contact Info

Product

Resources

About