Department
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Honors & Awards

Thomas E. Shenk

James A. Elkins Jr. Professor in the Life Sciences
Co-Director, Program in Global Health and Health Policy

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Phone (609) 258-5992
locationLewis Thomas Lab, 203
Phone Lab (609) 258-5993
Faculty Assistant
william mercado
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Phone (609) 258-1694

Research Focus

Human cytomegalovirus replication and pathogenesis

Cytomegaloviruses are members of the herpes virus family. Human cytomegalovirus (HCMV) infections are widespread and subclinical in the vast majority of cases, but the virus exhibits increased virulence in the very young and old and in immunocompromised individuals. Congenital infections cause life-long disabilities in a significant number of children. Transplant recipients, cancer patients, and AIDS patients, all of whom can exhibit decreased immune function, suffer a variety of clinical manifestations resulting from cytomegalovirus infection, including mononucleosis and pneumonia. There are also suggestions in the literature that HCMV might serve as a cofactor in certain cancers, atherosclerosis and immune senescence. The HCMV particle carries a viral genome comprised of linear double-stranded DNA that encodes more than 200 proteins, 23 microRNAs and a variety of additional non-coding RNAs. We study molecular mechanisms underlying HCMV replication and pathogenesis.

When the virus infects a fibroblast, epithelial cell or endothelial cell, it actively replicates and generates infectious progeny. We have studied the HCMV life cycle by using a mixture of genetic, biochemical, proteomic and metabolomic approaches. One area of special interest to us has been the identification of mechanisms by which the virus blocks defensive responses of the host cell, an area where the HCMV pUL38 protein plays a major role. We constructed a mutant virus unable to express pUL38 and found that it failed to efficiently express all viral genes tested and infected cells died of apoptosis before the viral replication cycle was completed. Using proteomic technology we discovered that pUL38 binds to the cellular tuberous sclerosis protein complex (TSC). This is a tumor suppressor complex that interprets stress signals and modulates the activity of mTOR, which in turn controls translation, fatty acid synthesis and many other vital processes in the cell. The pUL38 protein blocks the ability of TSC to respond to upstream signals, allowing the virus to continue to utilize cellular biosynthetic systems in spite of inducing a strong stress response. We have further probed the HCMV-host cell interaction by studying how metabolism is altered by infection. This work has revealed that HCMV induces glycolysis, nucleotide synthesis, citric acid cycle flux and lipid biosynthesis. Inhibition of the committed step of fatty acid synthesis and elongation, acetyl-CoA carboxylase, blocks HCMV replication, so we performed an siRNA screen and identified a substantial number of enzymes that sponsor fatty acid and lipid biosynthesis and are needed for successful virus replication. We are now working to elucidate how these enzymes are regulated, and one approach we have employed is to knock down each of the cell-coded kinases and test how the loss of function influences HCMV replication. This approach has identified several kinases that modulate the cell metabolome following infection. Most recently, we discovered that each of the seven human sirtuins is an HCMV restriction factor. Sirtuins are NAD+-dependent protein deacetylases, and we have begun to identify cellular targets of sirtuins that are critical for their anti-viral activity.

When HCMV infects a bone marrow stem cell or a monocyte, it doesn't replicate. Rather, it enters a state of quiescence termed latency. We have developed and validated two cell culture models for latency. Using these models, we have discovered that the virus transiently expresses a large number of viral products after infecting these cells, but after several days the viral genome becomes quiescent. A small number of viral proteins that continue to be expressed, including pUL138, a protein whose role in latency was first identified in our latency models. Others have shown that it controls the expression of at least one cell surface protein, and we are now following this clue by characterizing the cell surface proteome of uninfected monocytes as compared to monocytes harbouring latent HCMV. Our early results show that they are remarkably different!


Selected Publications

O'Connor C, DiMaggio PA Jr, Shenk T, Garcia BA. (2014) Quantitative proteomic discovery of dynamic epigenome changes that control human cytomegalovirus infection. Mol Cell Proteomics. Jul 1. [Epub ahead of print]

Casadevall A, Dermody TS, Imperiale MJ, Sandri-Goldin RM, Shenk T. (2014) On the need for a national board to assess dual use research of concern. J Virol. 88: 6535-37.  Pubmed

Dermody TS, Sandri-Goldin RM, Shenk T. (2014) Sequence changes associated with respiratory transmission of H7N1 influenza virus in mammals. J Virol. 88: 6533-34. Pubmed

Sharon-Friling R, Shenk T. (2014) Human cytomegalovirus pUL37x1-induced calcium flux activates PKCα, inducing altered cell shape and accumulation of cytoplasmic vesicles. Proc Natl Acad Sci. 111: E1140-48. Pubmed

Grady SL, Purdy JG, Rabinowitz JD, Shenk T. (2013) Argininosuccinate synthetase 1 depletion produces a metabolic state conducive to herpes simplex virus 1 infection. Proc Natl Acad Sci. 110: E5006-15. Pubmed

Koyuncu E, Purdy JG, Rabinowitz JD, Shenk T. (2013) Saturated very long chain Fatty acids are required for the production of infectious human cytomegalovirus progeny. PLoS Pathog. 9: e1003333. Pubmed

O'Connor CM, Shenk T. (2012) Human cytomegalovirus pUL78 G protein-coupled receptor homologue is required for timely cell entry in epithelial cells but not fibroblasts. J Virol. 86: 11425-33. Pubmed

Gudleski-O'Regan N, Greco TM, Cristea IM, Shenk T. (2012) Increased expression of LDL receptor-related protein 1 during human cytomegalovirus infection reduces virion cholesterol and infectivity. Cell Host Microbe. 12: 86-96. Pubmed

Grady SL, Hwang J, Vastag L, Rabinowitz JD, Shenk T. (2012) Herpes simplex virus type 1 infection activates poly(ADP-ribose) polymerase and triggers the degradation of poly(ADP-ribose) glycohydrolase. J Virol. 86: 8259-68. Pubmed

Terry LJ, Vastag L, Rabinowitz JD, Shenk T. (2012) Human kinome profiling identifies a requirement for AMP-activated protein kinase during human cytomegalovirus infection. Proc Natl Acad Sci. 109: 3071-76. PubMed

Liu ST, Sharon-Friling R, Ivanova P,...Shenk T. (2011) Synaptic vesicle-like lipidome of human cytomegalovirus virions reveals a role for SNARE machinery in virion egress. Proc Natl Acad Sci. 108: 12869-74. PubMed

Vastag L, Koyuncu E, Grady SL, Shenk TE, Rabinowitz JD. (2011) Divergent effects of human cytomegalovirus and herpes simplex virus-1 on cellular metabolism. PLoS Pathog. 7: e1002124. PubMed

O'Connor CM, Shenk T. (2011) Human cytomegalovirus pUS27 G protein-coupled receptor homologue is required for efficient spread by the extracellular route but not for direct cell-to-cell spread. J Virol. 85: 3700-07. PubMed

Womack A, Shenk T. (2010) Human cytomegalovirus tegument protein pUL71 is required for efficient virion egress. MBio. 1 pii: e00282-10. PubMed

Hargett D, Shenk TE. (2010) Experimental human cytomegalovirus latency in CD14+ monocytes. Proc Natl Acad Sci. 107: 20039-44. PubMed

Moorman NJ, Sharon-Friling R, Shenk T, Cristea IM. (2010) A targeted spatial-temporal proteomics approach implicates multiple cellular trafficking pathways in human cytomegalovirus virion maturation. Mol Cell Proteomics. 9: 851-60. PubMed

Terhune SS, Moorman NJ, Cristea IM,...Shenk T. (2010) Human cytomegalovirus UL29/28 protein interacts with components of the NuRD complex which promote accumulation of immediate-early RNA. PLoS Pathog. 6: e1000965. PubMed

Cristea IM, Moorman NJ, Terhune SS,....Shenk T. (2010) Human cytomegalovirus pUL83 stimulates activity of the viral immediate-early promoter through its interaction with the cellular IFI16 protein. J Virol. 84: 7803-14. PubMed

Moorman NJ, Shenk T. (2010) Rapamycin-resistant mTORC1 activity is required for herpesvirus replication. J Virol. 84: 5260-69. PubMed

Aoyagi M, Gaspar M, Shenk TE. (2010) Human cytomegalovirus UL69 protein facilitates translation by associating with the mRNA cap-binding complex and excluding 4EBP1. Proc Natl Acad Sci. 107: 2640-45. PubMed

Moorman NJ, Sharon-Friling R, Shenk T, Cristea IM. (2009) A targeted spatial-temporal proteomic approach implicates multiple cellular trafficking pathways in human cytomegalovirus virion maturation. Mol Cell Proteomics. 9: 851-60. PubMed

Mitchell DP, Savaryn JP, Moorman NJ, Shenk T, Terhune SS. (2009) Human cytomegalovirus UL28 and UL29 open reading frames encode a spliced mRNA and stimulate accumulation of immediate-early RNAs. J Virol. 83: 10187-97. PubMed

Lilja AE, Shenk T. (2008) Efficient replication of rhesus cytomegalovirus variants in multiple rhesus and human cell types. Proc Natl Acad Sci. 105: 19950-55. PubMed

Schröer J, Shenk T. (2008) Inhibition of cyclooxygenase activity blocks cell-to-cell spread of human cytomegalovirus. Proc Natl Acad Sci. 105: 19468-73. PubMed

Munger J, Bennett BD, Parikh A,...Shenk T, Rabinowitz JD. (2008) Systems-level metabolic flux profiling identifies fatty acid synthesis as a target for antiviral therapy. Nat Biotechnol. 26: 1179-86. PubMed

Cuevas-Bennett C, Shenk T. (2008) Dynamic histone H3 acetylation and methylation at human cytomegalovirus promoters during replication in fibroblasts. J Virol. 82: 9525-36. PubMed

Murphy E, Shenk T. (2008) Human cytomegalovirus genome. Curr Top Microbiol Immunol. 325: 1-19. PubMed

Moorman NJ, Cristea IM, Terhune SS, Rout MP, Chait BT, Shenk T. (2008) Human cytomegalovirus protein UL38 inhibits host cell stress responses by antagonizing the tuberous sclerosis protein complex. Cell Host Microbe. 3: 253-62. PubMed

Lilja AE, William Chang WL, Barry PA, Becerra SP, Shenk TE. (2007) Functional genetic analysis of Rhesus cytomegalovirus: Rh01 is an epithelial cell tropism factor. J Virol. 82: 2170-81. PubMed

Wang D, Yu QC, Schröer J, Murphy E, Shenk T. (2007) Human cytomegalovirus uses two distinct pathways to enter retinal pigmented epithelial cells. Proc Natl Acad Sci. 104: 20037-42. PubMed

Terhune S, Torigoi E, Moorman N,...Shenk T, Yu D (2007). Human Cytomegalovirus UL38 protein blocks apoptosis. J Virol. 81: 3109-23. PubMed

Weinberger LS, Shenk T. (2006) An HIV feedback resistor: auto-regulatory circuit deactivator and noise buffer. PLoS Biol. 5: e9. PubMed

Sharon-Friling R, Goodhouse J, Colberg-Poley AM, Shenk T. (2006) Human cytomegalovirus pUL37x1 induces the release of endoplasmic reticulum calcium stores. Proc Natl Acad Sci. 103: 19117-22. PubMed

Kulesza CA, Shenk T. (2006) Murine cytomegalovirus encodes a stable intron that facilitates persistent replication in the mouse. Proc Natl Acad Sci. 103: 18302-07. PubMed

Feng X, Schroer J, Yu D, Shenk T. (2006) Human Cytomegalovirus pUS24 is a virion protein that functions very early in the replication cycle. J Virol. 80: 8371-78. PubMed

Munger J, Yu D, Shenk T. (2006) UL26-deficient human cytomegalovirus produces virions with hypophosphorylated pp28 tegument protein that is unstable within newly infected cells. J Virol. 80: 3541-48. PubMed

Gaspar M, Shenk T. (2006) Human cytomegalovirus inhibits a DNA damage response by mislocalizing checkpoint proteins. Proc Natl Acad Sci. 103: 2821-26. PubMed

Wang D, Shenk T. (2005) Human cytomegalovirus virion protein complex required for epithelial and endothelial cell tropism. Proc Natl Acad Sci. 102: 18153-58. PubMed

Wang D, Shenk T. (2005) Human cytomegalovirus UL131 open reading frame is required for epithelial cell tropism. J Virol. 79: 10330-38. PubMed

Silva MC, Schroer J, Shenk T. (2005) Human cytomegalovirus cell-to-cell spread in the absence of an essential assembly protein. Proc Natl Acad Sci. 102: 2081-86. PubMed

Wang D, Bresnahan W, Shenk T. (2004) Human cytomegalovirus encodes a highly specific RANTES decoy receptor. Proc Natl Acad Sci. 101: 16642-47. PubMed

Varnum SM, Streblow DN, Monroe ME,...Shenk T,...Nelson JA. (2004) Identification of proteins in human cytomegalovirus (HCMV) particles: the HCMV proteome. J Virol. 78: 10960-66. PubMed

Terhune SS, Schroer J, Shenk T. (2004) RNAs are packaged into human cytomegalovirus virions in proportion to their intracellular concentration. J Virol. 78: 10390-98. PubMed

Nevels M, Paulus C, Shenk T. (2004) Human cytomegalovirus immediate-early 1 protein facilitates viral replication by antagonizing histone deacetylation. Proc Natl Acad Sci. 101: 17234-39. PubMed

Nevels M, Brune W, Shenk T. (2004) SUMOylation of the human cytomegalovirus 72-kilodalton IE1 protein facilitates expression of the 86-kilodalton IE2 protein and promotes viral replication. J Virol. 78: 7803-12. PubMed

Kulesza CA, Shenk T. (2004) Human cytomegalovirus 5-kilobase immediate-early RNA is a stable intron. J Virol. 78: 13182-89. PubMed

Goodrum F, Jordan CT, Terhune SS, High K, Shenk T. (2004) Differential outcomes of human cytomegalovirus infection in primitive hematopoietic cell subpopulations. Blood. 104: 687-95. PubMed

Challacombe JF, Rechtsteiner A, Gottardo R,...Shenk T, Altherr MR, Brettin TS (2004) Evaluation of the host transcriptional response to human cytomegalovirus infection. Physiol Genomics. 18: 51-62. PubMed

Adamo JE, Schroer J, Shenk T. (2004) Human cytomegalovirus TRS1 protein is required for efficient assembly of DNA-containing capsids. J Virol. 78: 10221-29. PubMed


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