Expression of the Drosophila melanogaster limk1 gene 3′-UTRs mRNA in yeast Saccharomyces cerevisiae
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The stability of mRNA and its translation efficacy in higher eukaryotes are influenced by the interaction of 3′-untranscribed regions (3′-UTRs) with microRNAs and RNA-binding proteins. Since Saccharomyces cerevisiae lack microRNAs, it is possible to evaluate the contribution of only 3′-UTRs’ and RNA-binding proteins’ interaction in post-transcriptional regulation. For this, the post-transcriptional regulation of Drosophila limk1 gene encoding for the key enzyme of actin remodeling was studied in yeast. Analysis of limk1 mRNA 3′-UTRs revealed the potential sites of yeast transcriptional termination. Computer modeling demonstrated the possibility of secondary structure formation in limk1 mRNA 3′-UTRs. For an evaluation of the functional activity of Drosophila 3′-UTRs in yeast, the reporter gene PHO5 encoding for yeast acid phosphatase (AP) fused to different variants of Drosophila limk1 mRNA 3′-UTRs (513, 1075, 1554 bp) was used. Assessments of AP activity and RT-PCR demonstrated that Drosophila limk1 Gene 3′-UTRs were functionally active and recognized in yeast. Therefore, yeast might be used as an appropriate model system for studies of 3′-UTR’s role in post-transcriptional regulation.
KeywordsAcid Phosphatase Minimal Free Energy PHO5 Gene Transcription Termination Site Cofilin Phosphorylation
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