Gene Expression Pattern in Caco-2 Cells following Rotavirus Infection

ABSTRACT Rotaviruses are recognized as the leading cause of severe dehydrating diarrhea in infants and young children worldwide. Preventive and therapeutic strategies are urgently needed to fight this pathogen. In tissue culture and in vivo, rotavirus induces structural and functional alterations in the host cell. In order to better understand the molecular mechanisms involved in the events after rotavirus infection, we identified host cellular genes whose mRNA levels changed after infection. For this analysis, we used microarrays containing more than 38,000 human cDNAs to study the transcriptional response of the human intestinal cell line Caco-2 to rotavirus infection. We found that 508 genes were differentially regulated >2-fold at 16 h after rotavirus infection, and only one gene was similarly regulated at 1 h postinfection. Of these transcriptional changes, 73% corresponded to the upregulation of genes, with the majority of them occurring late, at 12 or more hours postinfection. Some of the regulated genes were classified according to known biological function and included genes encoding integral membrane proteins, interferon-regulated genes, transcriptional and translational regulators, and calcium metabolism-related genes. A new picture of global transcriptional regulation in the infected cell is presented and families of genes which may be involved in viral pathogenesis are discussed.

[1]  E. Méndez,et al.  Heat Shock Cognate Protein 70 Is Involved in Rotavirus Cell Entry , 2002, Journal of Virology.

[2]  Roger E Bumgarner,et al.  Global Impact of Influenza Virus on Cellular Pathways Is Mediated by both Replication-Dependent and -Independent Events , 2001, Journal of Virology.

[3]  R. Krug,et al.  Influenza B virus NS1 protein inhibits conjugation of the interferon (IFN)‐induced ubiquitin‐like ISG15 protein , 2001, The EMBO journal.

[4]  J. Burnside,et al.  Induction of Host Gene Expression following Infection of Chicken Embryo Fibroblasts with Oncogenic Marek's Disease Virus , 2001, Journal of Virology.

[5]  Susana López,et al.  Integrin αvβ3 mediates rotavirus cell entry , 2000 .

[6]  Jean-Philippe Brunet,et al.  Rotavirus Infection Induces Cytoskeleton Disorganization in Human Intestinal Epithelial Cells: Implication of an Increase in Intracellular Calcium Concentration , 2000, Journal of Virology.

[7]  A. García-Sastre,et al.  Activation of Interferon Regulatory Factor 3 Is Inhibited by the Influenza A Virus NS1 Protein , 2000, Journal of Virology.

[8]  A. Servin,et al.  Rotavirus-Induced Structural and Functional Alterations in Tight Junctions of Polarized Intestinal Caco-2 Cell Monolayers , 2000, Journal of Virology.

[9]  L. Laimins,et al.  Microarray Analysis Identifies Interferon-Inducible Genes and Stat-1 as Major Transcriptional Targets of Human Papillomavirus Type 31 , 2000, Journal of Virology.

[10]  G. Sen,et al.  Novel functions of interferon-induced proteins. , 2000, Seminars in cancer biology.

[11]  Jean-Philippe Brunet,et al.  Rotavirus Infection Induces an Increase in Intracellular Calcium Concentration in Human Intestinal Epithelial Cells: Role in Microvillar Actin Alteration , 2000, Journal of Virology.

[12]  Roger E Bumgarner,et al.  Large-scale monitoring of host cell gene expression during HIV-1 infection using cDNA microarrays. , 2000, Virology.

[13]  Y. Takada,et al.  Integrins α2β1 and α4β1 Can Mediate SA11 Rotavirus Attachment and Entry into Cells , 2000, Journal of Virology.

[14]  D. Hober,et al.  MxA protein in capillary blood of children with viral infections , 1999, Journal of medical virology.

[15]  R. Shaw,et al.  The epithelial cell response to rotavirus infection. , 1999, Journal of immunology.

[16]  R. Donato,et al.  Functional roles of S100 proteins, calcium-binding proteins of the EF-hand type. , 1999, Biochimica et biophysica acta.

[17]  D. Trono,et al.  Cell-surface expression of CD4 reduces HIV-1 infectivity by blocking Env incorporation in a Nef- and Vpu-inhibitable manner , 1999, Current Biology.

[18]  H. Greenberg,et al.  Lack of a role for type I and type II interferons in the resolution of rotavirus-induced diarrhea and infection in mice. , 1999, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[19]  D. Gendrel,et al.  Interferon‐alpha in viral and bacterial gastroenteritis: a comparison with C‐reactive protein and interleukin‐6 , 1999, Acta paediatrica.

[20]  F. Michelangeli,et al.  Characterization of a Membrane Calcium Pathway Induced by Rotavirus Infection in Cultured Cells , 1999, Journal of Virology.

[21]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Bellamy,et al.  BiP (GRP78) and Endoplasmin (GRP94) Are Induced following Rotavirus Infection and Bind Transiently to an Endoplasmic Reticulum-Localized Virion Component , 1998, Journal of Virology.

[23]  T. Gingeras,et al.  Cellular gene expression altered by human cytomegalovirus: global monitoring with oligonucleotide arrays. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Jean Cohen,et al.  Rotavirus RNA‐binding protein NSP3 interacts with eIF4GI and evicts the poly(A) binding protein from eIF4F , 1998, The EMBO journal.

[25]  G. Stark,et al.  How cells respond to interferons. , 1998, Annual review of biochemistry.

[26]  Jean-Philippe Brunet,et al.  Rotavirus Infection Reduces Sucrase-Isomaltase Expression in Human Intestinal Epithelial Cells by Perturbing Protein Targeting and Organization of Microvillar Cytoskeleton , 1998, Journal of Virology.

[27]  M. Katze,et al.  Molecular mechanisms of interferon resistance mediated by viral-directed inhibition of PKR, the interferon-induced protein kinase. , 1998, Pharmacology & therapeutics.

[28]  D. Hober,et al.  The MxA protein levels in whole blood lysates of patients with various viral infections. , 1998, Journal of virological methods.

[29]  S. Goodbourn,et al.  Human Herpesvirus 8 Encodes an Interferon Regulatory Factor (IRF) Homolog That Represses IRF-1-Mediated Transcription , 1998, Journal of Virology.

[30]  A. Servin,et al.  Rotavirus is released from the apical surface of cultured human intestinal cells through nonconventional vesicular transport that bypasses the Golgi apparatus , 1997, Journal of virology.

[31]  D. Bass,et al.  Interferon gamma and interleukin 1, but not interferon alfa, inhibit rotavirus entry into human intestinal cell lines , 1997, Gastroenterology.

[32]  M. Estes,et al.  The rotavirus enterotoxin NSP4 mobilizes intracellular calcium in human intestinal cells by stimulating phospholipase C-mediated inositol 1,4,5-trisphosphate production. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Mcghee,et al.  Evidence for CD8+ T-cell immunity to murine rotavirus in the absence of perforin, fas, and gamma interferon , 1997, Journal of virology.

[34]  B. Finlay,et al.  Enteropathogenic Escherichia coli markedly decreases the resting membrane potential of Caco-2 and HeLa human epithelial cells , 1996, Infection and immunity.

[35]  R. Ménard,et al.  Bacterial entry into epithelial cells: the paradigm of Shigella. , 1996, Trends in microbiology.

[36]  R. Glass,et al.  New Lessons for Rotavirus Vaccines , 1996, Science.

[37]  M. Estes,et al.  Age-Dependent Diarrhea Induced by a Rotaviral Nonstructural Glycoprotein , 1996, Science.

[38]  C. Auffray,et al.  The I.M.A.G.E. Consortium: an integrated molecular analysis of genomes and their expression. , 1996, Genomics.

[39]  P. Cossart,et al.  E-Cadherin Is the Receptor for Internalin, a Surface Protein Required for Entry of L. monocytogenes into Epithelial Cells , 1996, Cell.

[40]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[41]  F. Michelangeli,et al.  Selective depletion of stored calcium by thapsigargin blocks rotavirus maturation but not the cytopathic effect , 1995, Journal of virology.

[42]  M. Estes,et al.  The nonstructural glycoprotein of rotavirus affects intracellular calcium levels , 1994, Journal of virology.

[43]  C. Dupont,et al.  Rotavirus Induces a‐Interferon Release in Children with Gastroenteritis , 1993, Journal of pediatric gastroenterology and nutrition.

[44]  P. Lengyel,et al.  The interferon system. A bird's eye view of its biochemistry. , 1992, The Journal of biological chemistry.

[45]  M. Estes,et al.  Comparative growth of different rotavirus strains in differentiated cells (MA104, HepG2, and CaCo-2). , 1991, Virology.

[46]  B. Finlay,et al.  Symmetric infection of rotavirus on polarized human intestinal epithelial (Caco-2) cells , 1991, Journal of virology.

[47]  C. Samuel,et al.  Antiviral actions of interferon. Interferon-regulated cellular proteins and their surprisingly selective antiviral activities. , 1991, Virology.

[48]  M. Lizano,et al.  The amino-terminal half of rotavirus SA114fM VP4 protein contains a hemagglutination domain and primes for neutralizing antibodies to the virus , 1991, Journal of virology.

[49]  J. D. del Castillo,et al.  Rotavirus infection alters Na+ and K+ homeostasis in MA-104 cells. , 1991, The Journal of general virology.

[50]  Richards Ab,et al.  Treatment of rotavirus infection in neonate and weanling pigs using natural human interferon alpha. , 1990 .

[51]  K. Fukudome,et al.  Comparison of human, simian, and bovine rotaviruses for requirement of sialic acid in hemagglutination and cell adsorption. , 1989, Virology.

[52]  C. Birch,et al.  Synthesis of human rotavirus polypeptides in cell culture , 1988, Journal of medical virology.

[53]  D. Keljo,et al.  Characterization of binding of simian rotavirus SA-11 to cultured epithelial cells. , 1988, Journal of pediatric gastroenterology and nutrition.

[54]  M. Rousset,et al.  Sucrase‐isomaltase: A marker of foetal and malignant epithelial cells of the human colon , 1983, International journal of cancer.

[55]  T. Orfeo,et al.  One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. , 1977, Journal of the National Cancer Institute.

[56]  T. Vesikari,et al.  Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis. , 1973, Lancet.

[57]  David Botstein,et al.  The Stanford Microarray Database , 2001, Nucleic Acids Res..

[58]  D. Botstein,et al.  The transcriptional program in the response of human fibroblasts to serum. , 1999, Science.

[59]  A. B. Richards,et al.  Treatment of rotavirus infection in neonate and weanling pigs using natural human interferon alpha. , 1990, Molecular biotherapy.

[60]  P. Pastoret,et al.  Experimental rotavirus diarrhoea in colostrum-deprived newborn calves: assay of treatment by administration of bacterially produced human interferon (Hu-IFN alpha 2). , 1985, Annales de recherches veterinaires. Annals of veterinary research.

[61]  C. La Bonnardière,et al.  Interferon activity in rotavirus infected newborn calves. , 1981, Annales de recherches veterinaires. Annals of veterinary research.