Autonomous Translational Pausing is Required for XBP1u MRNA Recruitment to the ER Via the SRP Pathway

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2016 Sep 22

PMID 27651490

Citations 37

Authors

Satoshi Kanda

Kota Yanagitani

Yukiko Yokota

Yuta Esaki

Kenji Kohno

Affiliations

Soon will be listed here.

Abstract

Unconventional mRNA splicing on the endoplasmic reticulum (ER) membrane is the sole conserved mechanism in eukaryotes to transmit information regarding misfolded protein accumulation to the nucleus to activate the stress response. In metazoans, the unspliced form of X-box-binding protein 1 (XBP1u) mRNA is recruited to membranes as a ribosome nascent chain (RNC) complex for efficient splicing. We previously reported that both hydrophobic (HR2) and translational pausing regions of XBP1u are important for the recruitment of its own mRNA to membranes. However, its precise location and the molecular mechanism of translocation are unclear. We show that XBP1u-RNC is specifically recruited to the ER membrane in an HR2- and translational pausing-dependent manner by immunostaining, fluorescent recovery after photobleaching, and biochemical analyses. Notably, translational pausing during XBP1u synthesis is indispensable for the recognition of HR2 by the signal recognition particle (SRP), resulting in efficient ER-specific targeting of the complex, similar to secretory protein targeting to the ER. On the ER, the XBP1u nascent chain is transferred from the SRP to the translocon; however, it cannot pass through the translocon or insert into the membrane. Therefore, our results support a noncanonical mechanism by which mRNA substrates are recruited to the ER for unconventional splicing.

Citing Articles

Homeostasis control in health and disease by the unfolded protein response.

Acosta-Alvear D, Harnoss J, Walter P, Ashkenazi A Nat Rev Mol Cell Biol. 2024; 26(3):193-212.

PMID: 39501044 DOI: 10.1038/s41580-024-00794-0.

Hepatocyte-specific GDF15 overexpression improves high-fat diet-induced obesity and hepatic steatosis in mice via hepatic FGF21 induction.

Takeuchi K, Yamaguchi K, Takahashi Y, Yano K, Okishio S, Ishiba H Sci Rep. 2024; 14(1):23993.

PMID: 39402176 PMC: 11473698. DOI: 10.1038/s41598-024-75107-8.

TXNDC5 Plays a Crucial Role in Regulating Endoplasmic Reticulum Activity through Different ER Stress Signaling Pathways in Hepatic Cells.

Bidooki S, Barranquero C, Sanchez-Marco J, Martinez-Beamonte R, Rodriguez-Yoldi M, Navarro M Int J Mol Sci. 2024; 25(13).

PMID: 39000233 PMC: 11241358. DOI: 10.3390/ijms25137128.

Exploring the IRE1 interactome: From canonical signaling functions to unexpected roles.

Le Goupil S, Laprade H, Aubry M, Chevet E J Biol Chem. 2024; 300(4):107169.

PMID: 38494075 PMC: 11007444. DOI: 10.1016/j.jbc.2024.107169.

Phylogeny-linked occurrence of ribosome stalling on the mRNAs of Arabidopsis unfolded protein response factor bZIP60 orthologs in divergent plant species.

Imamichi T, Kusumoto N, Aoyama H, Takamatsu S, Honda Y, Muraoka S Nucleic Acids Res. 2024; 52(8):4276-4294.

PMID: 38366760 PMC: 11077094. DOI: 10.1093/nar/gkae101.

References

Yanagitani K, Kimata Y, Kadokura H, Kohno K . Translational pausing ensures membrane targeting and cytoplasmic splicing of XBP1u mRNA. Science. 2011; 331(6017):586-9. DOI: 10.1126/science.1197142. View

Jungnickel B, Rapoport T . A posttargeting signal sequence recognition event in the endoplasmic reticulum membrane. Cell. 1995; 82(2):261-70. DOI: 10.1016/0092-8674(95)90313-5. View

Kimata Y, Ishiwata-Kimata Y, Ito T, Hirata A, Suzuki T, Oikawa D . Two regulatory steps of ER-stress sensor Ire1 involving its cluster formation and interaction with unfolded proteins. J Cell Biol. 2007; 179(1):75-86. PMC: 2064738. DOI: 10.1083/jcb.200704166. View

Halic M, Becker T, Pool M, Spahn C, Grassucci R, Frank J . Structure of the signal recognition particle interacting with the elongation-arrested ribosome. Nature. 2004; 427(6977):808-14. DOI: 10.1038/nature02342. View

Jurkin J, Henkel T, Faerch Nielsen A, Minnich M, Popow J, Kaufmann T . The mammalian tRNA ligase complex mediates splicing of XBP1 mRNA and controls antibody secretion in plasma cells. EMBO J. 2014; 33(24):2922-36. PMC: 4282640. DOI: 10.15252/embj.201490332. View

Fujiki Y, Hubbard A, Fowler S, Lazarow P . Isolation of intracellular membranes by means of sodium carbonate treatment: application to endoplasmic reticulum. J Cell Biol. 1982; 93(1):97-102. PMC: 2112113. DOI: 10.1083/jcb.93.1.97. View

High S, Flint N, Dobberstein B . Requirements for the membrane insertion of signal-anchor type proteins. J Cell Biol. 1991; 113(1):25-34. PMC: 2288911. DOI: 10.1083/jcb.113.1.25. View

Lakkaraju A, Mary C, Scherrer A, Johnson A, Strub K . SRP keeps polypeptides translocation-competent by slowing translation to match limiting ER-targeting sites. Cell. 2008; 133(3):440-51. PMC: 2430734. DOI: 10.1016/j.cell.2008.02.049. View

Lim B, Miyazaki R, Neher S, Siegele D, Ito K, Walter P . Heat shock transcription factor σ32 co-opts the signal recognition particle to regulate protein homeostasis in E. coli. PLoS Biol. 2013; 11(12):e1001735. PMC: 3866087. DOI: 10.1371/journal.pbio.1001735. View

10.

Akopian D, Shen K, Zhang X, Shan S . Signal recognition particle: an essential protein-targeting machine. Annu Rev Biochem. 2013; 82:693-721. PMC: 3805129. DOI: 10.1146/annurev-biochem-072711-164732. View

11.

Yoshida H, Matsui T, Yamamoto A, Okada T, Mori K . XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor. Cell. 2002; 107(7):881-91. DOI: 10.1016/s0092-8674(01)00611-0. View

12.

Kulak N, Pichler G, Paron I, Nagaraj N, Mann M . Minimal, encapsulated proteomic-sample processing applied to copy-number estimation in eukaryotic cells. Nat Methods. 2014; 11(3):319-24. DOI: 10.1038/nmeth.2834. View

13.

Pechmann S, Chartron J, Frydman J . Local slowdown of translation by nonoptimal codons promotes nascent-chain recognition by SRP in vivo. Nat Struct Mol Biol. 2014; 21(12):1100-5. PMC: 4488850. DOI: 10.1038/nsmb.2919. View

14.

Ron D, Walter P . Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol. 2007; 8(7):519-29. DOI: 10.1038/nrm2199. View

15.

Potter M, Nicchitta C . Endoplasmic reticulum-bound ribosomes reside in stable association with the translocon following termination of protein synthesis. J Biol Chem. 2002; 277(26):23314-20. DOI: 10.1074/jbc.M202559200. View

16.

Kosmaczewski S, Edwards T, Han S, Eckwahl M, Meyer B, Peach S . The RtcB RNA ligase is an essential component of the metazoan unfolded protein response. EMBO Rep. 2014; 15(12):1278-85. PMC: 4264930. DOI: 10.15252/embr.201439531. View

17.

Park E, Rapoport T . Mechanisms of Sec61/SecY-mediated protein translocation across membranes. Annu Rev Biophys. 2012; 41:21-40. DOI: 10.1146/annurev-biophys-050511-102312. View

18.

Walter P, Blobel G . Translocation of proteins across the endoplasmic reticulum III. Signal recognition protein (SRP) causes signal sequence-dependent and site-specific arrest of chain elongation that is released by microsomal membranes. J Cell Biol. 1981; 91(2 Pt 1):557-61. PMC: 2111983. DOI: 10.1083/jcb.91.2.557. View

19.

Keenan R, Freymann D, Stroud R, Walter P . The signal recognition particle. Annu Rev Biochem. 2001; 70:755-75. DOI: 10.1146/annurev.biochem.70.1.755. View

20.

Yanagitani K, Imagawa Y, Iwawaki T, Hosoda A, Saito M, Kimata Y . Cotranslational targeting of XBP1 protein to the membrane promotes cytoplasmic splicing of its own mRNA. Mol Cell. 2009; 34(2):191-200. DOI: 10.1016/j.molcel.2009.02.033. View