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Meq protein of Marek’s disease virus inhibits the transactivity of p53 |
Li Xiang-dong; Shen Yang; Qiu Ya-feng; Ma Zhi-yong |
Department of veterinary public health, Shanghai veterinary research institute, Chinese academy of agricultural sciences, Shanghai 200232, China |
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Abstract Objective To construct recombinant plasmids expressing wild type Meq (Meq-wt) and mutant Meq (Meq-mut) of Marek’s disease virus (MDV), and use the plasmids to explore the inhibitory activity of Meq gene on the transacitivty of p53. Methods Meq-wt was cloned by PCR, and Meq-mut, in which the p53 binding domain was deleted, was generated using PCR-based site-directed mutagenesis. The expressions of Meq-wt and -mut in primary chicken embryo fibroblasts (CEF) were detected by western blot. The p53-Luc report plasmid was co-tranfected with Meq-wt and -mut plasmids into CEF cells to evaluate the inhibitory activity of Meq gene on the transactivity of p53. The subcellular localizations of Meq proteins and p53 were also examined by immunofluorescent staining. Results DNA sequencing analysis showed that Meq-wt and –mut were obtained and inserted into the expression vector. The p53-Luc reporter gene assay showed that Meq inhibited the transactivity of p53. The Meq-wt protein was mostly localized in the nucleus of transfected cells and co-localized with p53 that is mainly localized in the nucleus, whereas the Meq-mut protein was distributed both in the cytoplasm and the nucleus. Conclusion Meq protein of MDV inhibits the transactivity of p53 and this inhibitory activity of Meq protein is exerted probably through directly binding to p53.
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Received: 01 January 1900
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Corresponding Authors:
Ma Zhi-yong
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