Different sequence elements are required for function of the cauliflower mosaic virus polyadenylation site in Saccharomyces cerevisiae compared with in plants.
We show that the polyadenylation site derived from the plant cauliflower mosaic virus (CaMV) is specifically functional in the yeast Saccharomyces cerevisiae. The mRNA 3' endpoints were mapped at the same position in yeast cells as in plants, and the CaMV polyadenylation site was recognized in an orientation-dependent manner. Mutational analysis of the CaMV 3'-end-formation signal revealed that multiple elements are essential for proper activity in yeast cells, including two upstream elements that are situated more than 100 and 43 to 51 nucleotides upstream of the poly(A) addition site and the sequences at or near the poly(A) addition site. A comparison of the sequence elements that are essential for proper function of the CaMV signal in yeast cells and plants showed that both organisms require a distal and a proximal upstream element but that these sequence elements are not identical in yeast cells and plants. The key element for functioning of the CaMV signal in yeast cells is the sequence TAGTATGTA, which is similar to a sequence previously proposed to act in yeast cells as a bipartite signal, namely, TAG ... TATGTA. Deletion of this sequence in the CaMV polyadenylation signal abolished 3'-end formation in yeast cells, and a single point mutation in this motif reduced the activity of the CaMV signal to below 15%. These results indicate that the bipartite sequence element acts as a signal for 3'-end formation in yeast cells but only together with other cis-acting elements.
Irniger
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Institute of Microbiology, Swiss Federal Institute of Technology, Zürich.
Sanfaçon
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Egli
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Braus
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D000444:Aldehyde Dehydrogenase / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D002872:Chromosome Deletion; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D010944:Plants / Q000235:genetics*; D010957:Plasmids; D011061:Poly A / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D012333:RNA, Messenger; D012367:RNA, Viral; D011061:Poly A; D012313:RNA; D000444:Aldehyde Dehydrogenase