A prevailing view is that the HSV-1 UL41 tegument protein mediates nonspecific degradation of both viral and cellular mRNAs by cleavage of the RNA at or near the 5' terminus and that viral gene expression prevails because of a higher rate of transcription of viral genes. Our studies that form the basis of this grant application challenge several aspects of this view. Specifically: (i) Microarray analyses revealed the up regulation of several hundred genes in infected cells as compared to mock-infected cells. Validation studies (Northern analyses, Real-Time PCR and immunoblots) revealed 4 groups of mRNAs. Group 1 exemplified by IEX-1, c-fos, cox-2 and Ikappabalpha mRNAs were indeed up regulated but the protein products were not made. These mRNAs were degraded in a UL41 dependent manner by deadenylation, endonucleolytic cleavage and 3' to 5' processive degradation. Moreover, the 5' domains of the partially degraded mRNAs tended to linger and were readily detected in cytoplasmic extracts. Group 2 exemplified by tristetraprolin (TTP) mRNA and group 3 exemplified by GADD45beta mRNA were also up regulated but were stable and indeed translated, again all in a UL41 dependent manner!! Actin mRNA, abundant but not up regulated was rapidly degraded. The RNAs forming groups 2 and 3 contain A-U rich elements characteristic of rapidly turning over mRNAs whereas actin and GADD45beta mRNAs do not have these elements. TTP is a stress-related protein involved in the degradation of A-U rich mRNAs by binding and translocating these mRNAs to exosomes. In essence, the degradation of mRNA mediated by the UL41 protein is not indiscriminant but highly selective. This application has 4 aims i.e.(1) To identify the sequences in TTP mRNA that confer selective stability to the mRNA in wild-type virus infected cells;.(2) To define the mechanism by which the TTP mRNA is spared from degradation in wild-type virus infected cells.(3) To define the basis for the differential rates of degradation of mRNAs containing A-U rich elements in wild-type virus infected cells as compared to uninfected cells or cells infected with ?UL41 mutant virus, and (4) to determine whether the numerous functions now associated with the UL41 protein are co-variant and the role of these functions in the biology of HSV-1.
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