Global view on the metabolism of RNA poly(A) tails in yeast Saccharomyces cerevisiae

The polyadenosine tail (poly[A]-tail) is a universal modification of eukaryotic messenger RNAs (mRNAs) and non-coding RNAs (ncRNAs). In budding yeast, Pap1-synthesized mRNA poly(A) tails enhance export and translation, whereas Trf4/5-mediated polyadenylation of ncRNAs facilitates degradation by the exosome. Using direct RNA sequencing, we decipher the extent of poly(A) tail dynamics in yeast defective in all relevant exonucleases, deadenylases, and poly(A) polymerases. Predominantly ncRNA poly(A) tails are 20-60 adenosines long. Poly(A) tails of newly transcribed mRNAs are 50 adenosine long on average, with an upper limit of 200. Exonucleolysis by Trf5-assisted nuclear exosome and cytoplasmic deadenylases trim the tails to 40 adenosines on average. Surprisingly, PAN2/3 and CCR4-NOT deadenylase complexes have a large pool of non-overlapping substrates mainly defined by expression level. Finally, we demonstrate that mRNA poly(A) tail length strongly responds to growth conditions, such as heat and nutrient deprivation. Nanopore direct-RNA libraries were prepared from cap-enriched murine mRNA, mixed with poly(A)+ S. cerevisiae RNA, with Direct RNA Sequencing Kit (ONT, SQK-RNA002). Sequencing was performed with MinION device (Flow cell type R9.4.1 RevC) and basecalled using Guppy (Oxford Nanopore Technologies).

Identifier
Source https://data.blue-cloud.org/search-details?step=~012C5C7F62606A61BA4A05A2AA214AD869E2B2464F7
Metadata Access https://data.blue-cloud.org/api/collections/C5C7F62606A61BA4A05A2AA214AD869E2B2464F7
Provenance
Instrument MinION; OXFORD_NANOPORE
Publisher Blue-Cloud Data Discovery & Access service; ELIXIR-ENA
Contributor INTERNATIONAL INSTITUTE OF MOLECULAR AND CELL BIOLOGY IN WARSAW
Publication Year 2024
OpenAccess true
Contact blue-cloud-support(at)maris.nl
Representation
Discipline Marine Science
Temporal Coverage Begin 2021-01-27T00:00:00Z
Temporal Coverage End 2021-07-29T00:00:00Z