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. 2002 Dec 24;99(26):17031-6.
doi: 10.1073/pnas.252588599. Epub 2002 Dec 12.

The patterns of accumulation of cellular RNAs in cells infected with a wild-type and a mutant herpes simplex virus 1 lacking the virion host shutoff gene

Brunella Taddeo  1 , Audrey Esclatine, Bernard Roizman
Affiliations

The patterns of accumulation of cellular RNAs in cells infected with a wild-type and a mutant herpes simplex virus 1 lacking the virion host shutoff gene

Brunella Taddeo et al. Proc Natl Acad Sci U S A. .

Abstract

Cellular RNA extracted from quiescent human foreskin fibroblasts harvested at 1, 3, 7, or 12 h after infection was profiled on Affymetrix HG-U95Av2 arrays designed to detect 12,626 unique human transcripts. We also profiled RNA extracted from cells harvested at 1 and 7 h after infection with a mutant lacking the gene (DeltaU(L)41) encoding a protein (vhs) brought into cells by the virus and responsible for nonselective degradation of RNA early in infection. We report the following: (i) of the 12 tested genes, up-regulated at least 3-fold relative to the values of mock infected cells, 9 were confirmed by real-time PCR. The microchip assays analyses indicate that there were 475 genes up-regulated > or =3-fold. The up-regulated genes were clustered into 15 groups with respect to temporal pattern of transcript accumulation, and classified into 20 groups on the basis of their function. The preponderance of cellular genes up-regulated early in infection play a predominant role in transcription, whereas those up-regulated at later times respond to intracellular stress or concern themselves with the cell cycle and apoptosis. (ii) The number of genes up-regulated early in infection was higher in cells infected with the DeltaU(L)41 mutant. Conversely, more genes were down-regulated late in infection with wild-type virus than with mutant viruses. Both observations are compatible with the known function of the U(L)41 gene product early in infection and with degradation of cellular RNAs in the absence of replenishment by de novo transcription of cellular genes.

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Figures

Fig 1.
Fig 1.
Total number of up-regulated genes in HSV-1(F)-infected cells according to fold increase relative to mock-infected cells. The total number of genes for each ratio value was calculated based on the distribution of genes reported in Table 1.
Fig 2.
Fig 2.
Real-time PCR analysis of selected genes whose expression was modulated in microarray profiling by HSV-1. Fold change (expressed as log ratio) observed by microarray analysis at the peak of induction and by real-time PCR at the same time for 16 selected genes. The dashed line corresponds to a 3-fold increase in the accumulated RNAs of infected cells compared with those of mock-infected cells.
Fig 3.
Fig 3.
Venn diagrams showing the distribution of differentially regulated genes during infection with HSV-1(F) or ΔUL41 mutant relative to those of mock-infected cells. (A) Up-regulated genes. (B) Down-regulated genes.
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