Universität zu Köln

Boehm, Volker ORCID: 0000-0001-7588-9842, Gerbracht, Jennifer V. ORCID: 0000-0002-3377-4649, Marx, Marie-Charlotte and Gehring, Niels H. ORCID: 0000-0001-7792-1164 (2016). Interrogating the degradation pathways of unstable mRNAs with XRN1-resistant sequences. Nat. Commun., 7. LONDON: NATURE PUBLISHING GROUP. ISSN 2041-1723

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Abstract

The turnover of messenger RNAs (mRNAs) is a key regulatory step of gene expression in eukaryotic cells. Due to the complexity of the mammalian degradation machinery, the contribution of decay factors to the directionality of mRNA decay is poorly understood. Here we characterize a molecular tool to interrogate mRNA turnover via the detection of XRN1-resistant decay fragments (xrFrag). Using nonsense-mediated mRNA decay (NMD) as a model pathway, we establish xrFrag analysis as a robust indicator of accelerated 50-30 mRNA decay. In tethering assays, monitoring xrFrag accumulation allows to distinguish decapping and endocleavage activities from deadenylation. Moreover, xrFrag analysis of mRNA degradation induced by miRNAs, AU-rich elements (AREs) as well as the 30 UTRs of cytokine mRNAs reveals the contribution of 50-30 decay and endonucleolytic cleavage. Our work uncovers formerly unrecognized modes of mRNA turnover and establishes xrFrag as a powerful tool for RNA decay analyses.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Boehm, Volker UNSPECIFIED orcid.org/0000-0001-7588-9842 UNSPECIFIED Gerbracht, Jennifer V. UNSPECIFIED orcid.org/0000-0002-3377-4649 UNSPECIFIED Marx, Marie-Charlotte UNSPECIFIED UNSPECIFIED UNSPECIFIED Gehring, Niels H. UNSPECIFIED orcid.org/0000-0001-7792-1164 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-252772
DOI:	10.1038/ncomms13691
Journal or Publication Title:	Nat. Commun.
Volume:	7
Date:	2016
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	LONDON
ISSN:	2041-1723
Language:	English
Faculty:	Faculty of Mathematics and Natural Sciences
Divisions:	Faculty of Mathematics and Natural Sciences > Department of Biology > Institute for Genetics
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language CCR4-NOT DEADENYLASE COMPLEX; STRUCTURAL BASIS; ENDONUCLEOLYTIC CLEAVAGE; SMG5-SMG7 HETERODIMER; MAMMALIAN-CELLS; NONCODING RNA; DECAY; SURVEILLANCE; TRANSCRIPTS; MECHANISMS Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/25277