Effects of sequence motifs in the yeast 3′ untranslated region determined from massively parallel assays of random sequences View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2021-10-18

AUTHORS

Andrew Savinov, Benjamin M. Brandsen, Brooke E. Angell, Josh T. Cuperus, Stanley Fields

ABSTRACT

BackgroundThe 3′ untranslated region (UTR) plays critical roles in determining the level of gene expression through effects on activities such as mRNA stability and translation. Functional elements within this region have largely been identified through analyses of native genes, which contain multiple co-evolved sequence features.ResultsTo explore the effects of 3′ UTR sequence elements outside of native sequence contexts, we analyze hundreds of thousands of random 50-mers inserted into the 3′ UTR of a reporter gene in the yeast Saccharomyces cerevisiae. We determine relative protein expression levels from the fitness of transformants in a growth selection. We find that the consensus 3′ UTR efficiency element significantly boosts expression, independent of sequence context; on the other hand, the consensus positioning element has only a small effect on expression. Some sequence motifs that are binding sites for Puf proteins substantially increase expression in the library, despite these proteins generally being associated with post-transcriptional downregulation of native mRNAs. Our measurements also allow a systematic examination of the effects of point mutations within efficiency element motifs across diverse sequence backgrounds. These mutational scans reveal the relative in vivo importance of individual bases in the efficiency element, which likely reflects their roles in binding the Hrp1 protein involved in cleavage and polyadenylation.ConclusionsThe regulatory effects of some 3′ UTR sequence features, like the efficiency element, are consistent regardless of sequence context. In contrast, the consequences of other 3′ UTR features appear to be strongly dependent on their evolved context within native genes. More... »

PAGES

293

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/s13059-021-02509-6

DOI

http://dx.doi.org/10.1186/s13059-021-02509-6

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1141971475

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/34663436


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