Transcriptome analysis of fibroblast cells immediate-early after human cytomegalovirus infection.
Human cytomegalovirus (HCMV) has been shown to have the potential to alter cellular gene expression early after infection. However, one-gene approaches and the use of closed system gene expression technologies have identified only few cellular genes whose activity changed immediate-early. We therefore used serial analysis of gene expression (SAGE) to investigate the transcriptional program of human fibroblasts in response to HCMV in the immediate-early phase of infection. Differential expression of various cellular genes was monitored. Transcriptional expression changes of genes coding for ribosomal proteins reflected a general cellular response to starvation and stress. But differential regulation of genes coding for transcription factors and proteins associated with cellular metabolism, homeostasis and cell structure may represent transcriptional alterations in response to HCMV infection. Expression kinetics by 5' nuclease fluorigenic real-time PCR of selected genes revealed partial protection of infected cells against initial stress-associated alterations of gene expression and indicated fluctuations of transcriptional levels over time. Additionally, agreement with the quantitative results obtained by SAGE was observed only for genes up-regulated in HCMV-infected cells. This finding pointed to various technical and statistical parameters that all may be critical for quantitative transcriptome studies using global approaches, especially when exploring biological systems in a critical phase of cellular physiology.
Kenzelmann
M
M
Institute of Medical Microbiology, University of Bern, Friedbühlstrasse 51, Bern, 3010, Switzerland. m.kenzelmann@dkfz.de
Mühlemann
K
K
D002453:Cell Cycle; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism* / Q000821:virology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D017781:Genes, Immediate-Early / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D016133:Polymerase Chain Reaction; D019914:Taq Polymerase / Q000378:metabolism; D014158:Transcription, Genetic
D019914:Taq Polymerase