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PMID: 10357823 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mechanism of non-spliceosomal mRNA splicing in the unfolded protein response pathway.

The EMBO journal ·Vol. 18 ·No. 11 ·1999-06-01 ·Pages 3119-32

Gonzalez TN, Sidrauski C, Dörfler S, Walter P

Abstract

The unfolded protein response is an intracellular signaling pathway that, in response to accumulation of misfolded proteins in the lumen of the endoplasmic reticulum (ER), upregulates transcription of ER resident chaperones. A key step in this pathway is the non-conventional, regulated splicing of the mRNA encoding the positive transcriptional regulator Hac1p. In the yeast Saccharomyces cerevisiae, the bifunctional transmembrane kinase/endoribonuclease Ire1p cleaves HAC1 mRNA at both splice junctions and tRNA ligase joins the two exons together. We have reconstituted HAC1 mRNA splicing in an efficient in vitro reaction and show that, in many ways, the mechanism of HAC1 mRNA splicing resembles that of pre-tRNA splicing. In particular, Ire1p endonucleolytic cleavage leaves 2', 3'-cyclic phosphates, the excised exons remain associated by base pairing, and exon ligation by tRNA ligase follows the same chemical steps as for pre-tRNA splicing. To date, this mechanism of RNA processing is unprecedented for a messenger RNA. In contrast to the striking similarities to tRNA splicing, the structural features of the splice junctions recognized by Ire1p differ from those recognized by tRNA endonuclease. We show that small stem-loop structures predicted to form at both splice junctions of HAC1 mRNA are required and sufficient for Ire1p cleavage.

MeSH Terms
Base Pairing Base Sequence Basic-Leucine Zipper Transcription Factors Exons/genetics Fungal Proteins/genetics,metabolism Guanosine/genetics,metabolism Hydrolysis Introns/genetics Membrane Glycoproteins/metabolism Molecular Sequence Data Mutation Nucleic Acid Conformation Nucleotides/genetics,metabolism Phosphates/metabolism Protein Folding Protein Serine-Threonine Kinases RNA Ligase (ATP)/metabolism RNA Splicing/genetics RNA, Messenger/chemistry,genetics,metabolism RNA, Transfer/chemistry,genetics,metabolism Regulatory Sequences, Nucleic Acid/genetics Repressor Proteins/genetics Saccharomyces cerevisiae/enzymology,genetics,metabolism Saccharomyces cerevisiae Proteins Spliceosomes/physiology Substrate Specificity Transcription Factors
Chemicals
Basic-Leucine Zipper Transcription Factors Fungal Proteins HAC1 protein, S cerevisiae Membrane Glycoproteins Nucleotides Phosphates RNA, Messenger Repressor Proteins Saccharomyces cerevisiae Proteins Transcription Factors Guanosine RNA, Transfer IRE1 protein, S cerevisiae Protein Serine-Threonine Kinases RNA Ligase (ATP)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Gonzalez T N
Howard Hughes Medical Institute and Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, CA 94143-0448, USA.
Sidrauski C
Dörfler S
Walter P
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/18.11.3119
Published
1999-06-01
Pages
3119-32
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171393
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/18.11.3119 PMC Full Text View journal details
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