Differential effects of polyadenylation regions on gene expression in mammalian cells.
The steady-state level attained for any protein in mammalian cells is in part determined by its steady-state level of mRNA. Sequence information in and around the 3' end of an RNA which is involved in specifying and regulating polyadenylation [poly(A)] may have important consequences on mRNA levels, and ultimately on expression of the protein product. In this report we compare the effects on gene expression which result from placing several different poly(A) regions, or no poly(A) region, downstream from a marker gene (galactokinase or galK) that can be readily assayed in mammalian cells. Our results demonstrate that the presence of a poly(A) region is important for efficient gene expression and that the use of the poly(A) region of bovine growth hormone (bGH) reproducibly results in three times higher expression than that of SV40 early or human collagen poly(A) regions. We further demonstrate that changing the promoter region on these chimeric transcription units does not change the effect of the poly(A) region. Neither does changing the assay gene, since comparison of the same poly(A) regions behind another marker gene (xanthine-guanine phosphoribosyl transferase or xgprt) leads to identical differences in expression. When we examine the levels of poly(A)+ RNA that result from each transcription unit, we find that they correlate precisely with the gene expression levels. Apparently the 3' end of an RNA is a determinant of steady-state mRNA levels and, in turn, the subsequent production of the protein product.
Pfarr
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Rieser
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Woychik
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Rottman
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Rosenberg
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Reff
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ME
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D002417:Cattle; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics; D006224:Cricetinae; D004274:DNA, Recombinant; D005686:Galactokinase / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000235:genetics; D006801:Humans; D010430:Pentosyltransferases / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D011061:Poly A / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D013045:Species Specificity
D001426:Bacterial Proteins; D004274:DNA, Recombinant; D012333:RNA, Messenger; D011994:Recombinant Proteins; D011061:Poly A; D013006:Growth Hormone; D003094:Collagen; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase; D005686:Galactokinase