Cargando…

Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae

Most functional eukaryotic mRNAs contain a 5′ 7-methylguanosine (m(7)G) cap. Although capping is essential for many biological processes including mRNA processing, export and translation, the fate of uncapped transcripts has not been studied extensively. Here, we employed fast nuclear depletion of t...

Descripción completa

Detalles Bibliográficos
Autores principales: Zanin, Onofrio, Eastham, Matthew, Winczura, Kinga, Ashe, Mark, Martinez-Nunez, Rocio T., Hebenstreit, Daniel, Grzechnik, Pawel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10622555/
https://www.ncbi.nlm.nih.gov/pubmed/37919390
http://dx.doi.org/10.1038/s42003-023-05495-6
_version_ 1785130565259755520
author Zanin, Onofrio
Eastham, Matthew
Winczura, Kinga
Ashe, Mark
Martinez-Nunez, Rocio T.
Hebenstreit, Daniel
Grzechnik, Pawel
author_facet Zanin, Onofrio
Eastham, Matthew
Winczura, Kinga
Ashe, Mark
Martinez-Nunez, Rocio T.
Hebenstreit, Daniel
Grzechnik, Pawel
author_sort Zanin, Onofrio
collection PubMed
description Most functional eukaryotic mRNAs contain a 5′ 7-methylguanosine (m(7)G) cap. Although capping is essential for many biological processes including mRNA processing, export and translation, the fate of uncapped transcripts has not been studied extensively. Here, we employed fast nuclear depletion of the capping enzymes in Saccharomyces cerevisiae to uncover the turnover of the transcripts that failed to be capped. We show that although the degradation of cap-deficient mRNA is dominant, the levels of hundreds of non-capped mRNAs increase upon depletion of the capping enzymes. Overall, the abundance of non-capped mRNAs is inversely correlated to the expression levels, altogether resembling the effects observed in cells lacking the cytoplasmic 5′−3′ exonuclease Xrn1 and indicating differential degradation fates of non-capped mRNAs. The inactivation of the nuclear 5′−3′ exonuclease Rat1 does not rescue the non-capped mRNA levels indicating that Rat1 is not involved in their degradation and consequently, the lack of the capping does not affect the distribution of RNA Polymerase II on the chromatin. Our data indicate that the cap presence is essential to initiate the Xrn1-dependent degradation of mRNAs underpinning the role of 5′ cap in the Xrn1-dependent buffering of the cellular mRNA levels.
format Online
Article
Text
id pubmed-10622555
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-106225552023-11-04 Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae Zanin, Onofrio Eastham, Matthew Winczura, Kinga Ashe, Mark Martinez-Nunez, Rocio T. Hebenstreit, Daniel Grzechnik, Pawel Commun Biol Article Most functional eukaryotic mRNAs contain a 5′ 7-methylguanosine (m(7)G) cap. Although capping is essential for many biological processes including mRNA processing, export and translation, the fate of uncapped transcripts has not been studied extensively. Here, we employed fast nuclear depletion of the capping enzymes in Saccharomyces cerevisiae to uncover the turnover of the transcripts that failed to be capped. We show that although the degradation of cap-deficient mRNA is dominant, the levels of hundreds of non-capped mRNAs increase upon depletion of the capping enzymes. Overall, the abundance of non-capped mRNAs is inversely correlated to the expression levels, altogether resembling the effects observed in cells lacking the cytoplasmic 5′−3′ exonuclease Xrn1 and indicating differential degradation fates of non-capped mRNAs. The inactivation of the nuclear 5′−3′ exonuclease Rat1 does not rescue the non-capped mRNA levels indicating that Rat1 is not involved in their degradation and consequently, the lack of the capping does not affect the distribution of RNA Polymerase II on the chromatin. Our data indicate that the cap presence is essential to initiate the Xrn1-dependent degradation of mRNAs underpinning the role of 5′ cap in the Xrn1-dependent buffering of the cellular mRNA levels. Nature Publishing Group UK 2023-11-02 /pmc/articles/PMC10622555/ /pubmed/37919390 http://dx.doi.org/10.1038/s42003-023-05495-6 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Zanin, Onofrio
Eastham, Matthew
Winczura, Kinga
Ashe, Mark
Martinez-Nunez, Rocio T.
Hebenstreit, Daniel
Grzechnik, Pawel
Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae
title Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae
title_full Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae
title_fullStr Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae
title_full_unstemmed Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae
title_short Ceg1 depletion reveals mechanisms governing degradation of non-capped RNAs in Saccharomyces cerevisiae
title_sort ceg1 depletion reveals mechanisms governing degradation of non-capped rnas in saccharomyces cerevisiae
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10622555/
https://www.ncbi.nlm.nih.gov/pubmed/37919390
http://dx.doi.org/10.1038/s42003-023-05495-6
work_keys_str_mv AT zaninonofrio ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae
AT easthammatthew ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae
AT winczurakinga ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae
AT ashemark ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae
AT martineznunezrociot ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae
AT hebenstreitdaniel ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae
AT grzechnikpawel ceg1depletionrevealsmechanismsgoverningdegradationofnoncappedrnasinsaccharomycescerevisiae