Effects of splitting alternative KlCYC1 3'-UTR regions on processing: metabolic consequences and biotechnological applications.
To analyze the functionality of alternative 3'-UTR processing in the yeast Kluyveromyces lactis, recombinant forms of the KlCYC1 gene containing the proximal (1-713) or the distal (699-1194) 3'-UTR region (positions related to the TAA stop codon) were obtained. The cells expressing the gene with proximal 3'-UTR showed the same growth phenotype as the wild type. When the gene expressed only the distal region, a single transcript was generated and its expression was increased in late-growth phases. Cells expressing the alternative distal 3'-UTR region showed differences in their levels of cytochrome c biomass and ethanol production with respect to the wild type. The split 3'-UTR regions were also functional as separate processing units in Saccharomyces cerevisiae. The importance of our results in recombinant gene expression applications will be discussed.
Seoane
Silvia
S
Area de Bioquímica, Dpto de Bioloxía Celular e Molecular, Facultad de Ciencias, Campus da Zapateira S/N, 15071 A Coruña, Spain.
Guiard
Bernard
B
Rodríguez-Torres
Ana-María
AM
Freire-Picos
María Angeles
MA
D020413:3' Untranslated Regions; D045304:Cytochromes c / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D007716:Kluyveromyces / Q000235:genetics*; D026723:Polyadenylation / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics
D020413:3' Untranslated Regions; D005656:Fungal Proteins; D045304:Cytochromes c