Analysis of cis-regulatory elements in the 5' untranslated region of murine leukemia virus controlling protein expression.
It has previously been reported by us that high-level expression of the Env protein of Fr-MLV clone A8 in brains is crucial for induction of spongiform neurodegeneration, and that the 0.3-kb fragment containing the R, U5, and the 5' leader sequence of A8 is responsible for neuropathogenicity. In the present study, the role of the 5' untranslated region in protein expression was investigated. Luciferase expression vectors containing the LTR (R-U3-U5) and 5' leader sequence of A8 and non-neuropathogenic 57 Fr-MLV, designated gl-A8 and gl-57, and their chimeric vectors, were constructed, and transfected into rat glial cells F10. Replacement of the region containing the 3' half of R, U5, and 5' leader sequence of gl-A8 with that of 57 showed a reduction in luciferase activities, and replacement of this region of gl-57 with that of A8 showed increased luciferase activity. These results show that the region containing the 3' half of R, U5, and 5' leader sequence of A8 more efficiently up-regulates protein expression than 57. In particular, the 3' half of 5' leader of A8 was most responsible for the up-regulation of protein expression. Of interest, after replacement of the fragments between A8 and 57, changes in the activities of vectors containing A8-U3 paralleled the amount of mRNA, but the activities of vectors containing 57-U3 did not. Furthermore, it is suggested that the region containing R, U5, and the 5' leader sequence influences transcriptional or post-transcriptional steps, depending on the upstream sequence containing enhancer elements and promoter.
Yamamoto
Naoki
N
Department of Bioinformatics, Faculty of Engineering, Soka University, Hachioji, Tokyo 192-8577, Japan.
Takase-Yoden
Sayaka
S
D020121:5' Untranslated Regions; D000818:Animals; D019677:Artificial Gene Fusion; D001483:Base Sequence; D002460:Cell Line; D017930:Genes, Reporter; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009457:Neuroglia / Q000821:virology; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000096:biosynthesis; D012367:RNA, Viral / Q000235:genetics*; D051381:Rats; D011995:Recombination, Genetic; D050436:Regulatory Elements, Transcriptional; D014764:Viral Proteins / Q000096:biosynthesis*
D020121:5' Untranslated Regions; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D008156:Luciferases