Murine Gammaherpesvirus-68 Encodes Homologues of Thymidine Kinase and Glycoprotein H: Sequence, Expression, and Characterization of Pyrimidine Kinase Activity

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• 作者：PEPPER ; S. DE V. ; STEWART ; J. P. ; ARRAND ; J. R. ; MACKETT ; M.
• 刊名：Virology
• 出版年：1996
• 期刊代码：108_00426822
• 类别：imm
• 出版时间：May 15, 1996
• 卷：219
• 期：2
• 页码：475-479
• 文件大小：161 K

摘要

We have sequenced a 4.5-kb fragment of DNA spanning the junction of theBamHI D and E fragments of murine gammaherpesvirus-68 (MHV-68). This sequence was found to code for two major open reading frames (orfs) of 1934 and 2192 bp which showed significant homology to the thymidine kinase (TK) and glycoprotein H (gH) sequences of other gammaherpesviruses. Upstream from the TK gene another orf was found which showed amino acid sequence homology to the HSV1 UL24 gene. Analysis of the 1934-bp orf revealed the presence of all six of the recognized sites that are conserved between herpesvirus TKs although, uniquely among sequenced herpesvirus TK enzymes, MHV-68 lacks the consensus nucleotide binding site GXXGXGK, the second glycine being replaced by alanine. The MHV-68 TK has a predictedM_rof 68,443, while the gH is predicted to have aM_rof 82,890. Northern blot analysis showed an early TK message of 2.6 kb and a late gH-specific message of 2.5 kb. Both TK and gH probes detected a 4.3-kb late message, implying that this late message spans gH and TK. The TK coding sequence was expressed using anin vitrotranscription translation system and was shown to encode functional TK activity.