Cargando…
Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells
Post-transcriptional 3′ end addition of nucleotides is important in a variety of RNA decay pathways. We have examined the 3′ end addition of nucleotides during the decay of the Hsp70 mRNA and a corresponding reporter RNA in Drosophila S2 cells by conventional sequencing of cDNAs obtained after mRNA...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4748820/ https://www.ncbi.nlm.nih.gov/pubmed/26786835 http://dx.doi.org/10.1261/rna.053942.115 |
_version_ | 1782415186691358720 |
---|---|
author | Harnisch, Christiane Cuzic-Feltens, Simona Dohm, Juliane C. Götze, Michael Himmelbauer, Heinz Wahle, Elmar |
author_facet | Harnisch, Christiane Cuzic-Feltens, Simona Dohm, Juliane C. Götze, Michael Himmelbauer, Heinz Wahle, Elmar |
author_sort | Harnisch, Christiane |
collection | PubMed |
description | Post-transcriptional 3′ end addition of nucleotides is important in a variety of RNA decay pathways. We have examined the 3′ end addition of nucleotides during the decay of the Hsp70 mRNA and a corresponding reporter RNA in Drosophila S2 cells by conventional sequencing of cDNAs obtained after mRNA circularization and by deep sequencing of dedicated libraries enriched for 3′ decay intermediates along the length of the mRNA. Approximately 5%–10% of 3′ decay intermediates carried nonencoded oligo(A) tails with a mean length of 2–3 nucleotides. RNAi experiments showed that the oligoadenylated RNA fragments were intermediates of exosomal decay and the noncanonical poly(A) polymerase Trf4-1 was mainly responsible for A addition. A hot spot of A addition corresponded to an intermediate of 3′ decay that accumulated upon inhibition of decapping, and knockdown of Trf4-1 increased the abundance of this intermediate, suggesting that oligoadenylation facilitates 3′ decay. Oligoadenylated 3′ decay intermediates were found in the cytoplasmic fraction in association with ribosomes, and fluorescence microscopy revealed a cytoplasmic localization of Trf4-1. Thus, oligoadenylation enhances exosomal mRNA degradation in the cytoplasm. |
format | Online Article Text |
id | pubmed-4748820 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-47488202017-03-01 Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells Harnisch, Christiane Cuzic-Feltens, Simona Dohm, Juliane C. Götze, Michael Himmelbauer, Heinz Wahle, Elmar RNA Article Post-transcriptional 3′ end addition of nucleotides is important in a variety of RNA decay pathways. We have examined the 3′ end addition of nucleotides during the decay of the Hsp70 mRNA and a corresponding reporter RNA in Drosophila S2 cells by conventional sequencing of cDNAs obtained after mRNA circularization and by deep sequencing of dedicated libraries enriched for 3′ decay intermediates along the length of the mRNA. Approximately 5%–10% of 3′ decay intermediates carried nonencoded oligo(A) tails with a mean length of 2–3 nucleotides. RNAi experiments showed that the oligoadenylated RNA fragments were intermediates of exosomal decay and the noncanonical poly(A) polymerase Trf4-1 was mainly responsible for A addition. A hot spot of A addition corresponded to an intermediate of 3′ decay that accumulated upon inhibition of decapping, and knockdown of Trf4-1 increased the abundance of this intermediate, suggesting that oligoadenylation facilitates 3′ decay. Oligoadenylated 3′ decay intermediates were found in the cytoplasmic fraction in association with ribosomes, and fluorescence microscopy revealed a cytoplasmic localization of Trf4-1. Thus, oligoadenylation enhances exosomal mRNA degradation in the cytoplasm. Cold Spring Harbor Laboratory Press 2016-03 /pmc/articles/PMC4748820/ /pubmed/26786835 http://dx.doi.org/10.1261/rna.053942.115 Text en © 2016 Harnisch et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Article Harnisch, Christiane Cuzic-Feltens, Simona Dohm, Juliane C. Götze, Michael Himmelbauer, Heinz Wahle, Elmar Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells |
title | Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells |
title_full | Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells |
title_fullStr | Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells |
title_full_unstemmed | Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells |
title_short | Oligoadenylation of 3′ decay intermediates promotes cytoplasmic mRNA degradation in Drosophila cells |
title_sort | oligoadenylation of 3′ decay intermediates promotes cytoplasmic mrna degradation in drosophila cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4748820/ https://www.ncbi.nlm.nih.gov/pubmed/26786835 http://dx.doi.org/10.1261/rna.053942.115 |
work_keys_str_mv | AT harnischchristiane oligoadenylationof3decayintermediatespromotescytoplasmicmrnadegradationindrosophilacells AT cuzicfeltenssimona oligoadenylationof3decayintermediatespromotescytoplasmicmrnadegradationindrosophilacells AT dohmjulianec oligoadenylationof3decayintermediatespromotescytoplasmicmrnadegradationindrosophilacells AT gotzemichael oligoadenylationof3decayintermediatespromotescytoplasmicmrnadegradationindrosophilacells AT himmelbauerheinz oligoadenylationof3decayintermediatespromotescytoplasmicmrnadegradationindrosophilacells AT wahleelmar oligoadenylationof3decayintermediatespromotescytoplasmicmrnadegradationindrosophilacells |