Abstract
Friend murine leukemia virus (Fr-MLV) produces unspliced mRNAs, as well as singly-spliced mRNAs by which the Env protein is generated. We investigated the role of the intron within the Fr-MLV gene in Env expression using vectors with serially truncated introns. The up-regulatory regions, the 361–878 and the 5135–5399 fragments, and the down-regulatory regions, the 1904–2849 and the 3995–4287 fragments, were found to influence the splicing efficiency. The Env protein expression level was proportional to the amount of env-mRNA. Furthermore, we found that the splicing process is important to acquire env-mRNA stability, and it also promotes translation of the Env protein. These splicing-dependent phenomena were not observed when luciferase expression vectors were analyzed. These findings indicated that the Env protein expression level in MLV is regulated at multiple steps: the env-mRNA expression level is determined by the splicing efficiency, and the stability of env-mRNA is acquired through the splicing process.
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