Structural and mutational analysis of the ribosome-arresting human XBP1u.
Amino Acid Sequence
Animals
Biomechanical Phenomena
DNA Mutational Analysis
Endoribonucleases
/ metabolism
Humans
Models, Molecular
Mutagenesis
Peptides
/ chemistry
Peptidyl Transferases
/ metabolism
Protein Binding
Protein Serine-Threonine Kinases
/ metabolism
Protein Stability
Rabbits
Ribosomes
/ metabolism
SEC Translocation Channels
/ chemistry
Signal Recognition Particle
/ metabolism
Signal Transduction
Unfolded Protein Response
X-Box Binding Protein 1
/ chemistry
XBP1
biochemistry
chemical biology
molecular biophysics
ribosome
structural biology
translational pausing
Journal
eLife
ISSN: 2050-084X
Titre abrégé: Elife
Pays: England
ID NLM: 101579614
Informations de publication
Date de publication:
27 06 2019
27 06 2019
Historique:
received:
20
02
2019
accepted:
20
06
2019
pubmed:
28
6
2019
medline:
20
2
2020
entrez:
28
6
2019
Statut:
epublish
Résumé
XBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM structure of the stalled human XBP1u AP. It forms a unique turn in the ribosomal exit tunnel proximal to the peptidyl transferase center where it causes a subtle distortion, thereby explaining the temporary translational arrest induced by XBP1u. During ribosomal pausing the hydrophobic region 2 (HR2) of XBP1u is recognized by SRP, but fails to efficiently gate the Sec61 translocon. An exhaustive mutagenesis scan of the XBP1u AP revealed that only 8 out of 20 mutagenized positions are optimal; in the remaining 12 positions, we identify 55 different mutations increase the level of translational arrest. Thus, the wildtype XBP1u AP induces only an intermediate level of translational arrest, allowing efficient targeting by SRP without activating the Sec61 channel.
Identifiants
pubmed: 31246176
doi: 10.7554/eLife.46267
pii: 46267
pmc: PMC6624018
doi:
pii:
Substances chimiques
Peptides
0
SEC Translocation Channels
0
Signal Recognition Particle
0
X-Box Binding Protein 1
0
Peptidyl Transferases
EC 2.3.2.12
ERN1 protein, human
EC 2.7.11.1
Protein Serine-Threonine Kinases
EC 2.7.11.1
Endoribonucleases
EC 3.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Deutsche Forschungsgemeinschaft
ID : QBM (Quantitative Biosciences Munich) Graduate School Fellowship
Pays : International
Organisme : Knut och Alice Wallenbergs Stiftelse
ID : 2012.0282
Pays : International
Organisme : Vetenskapsrådet
ID : 621-2014-3713
Pays : International
Organisme : Cancerfonden
ID : 15 0888
Pays : International
Organisme : Japan Society for the Promotion of Science
ID : JP26116006
Pays : International
Organisme : Deutsche Forschungsgemeinschaft
ID : GRK1721
Pays : International
Organisme : Boehringer Ingelheim Fonds
ID : Graduate Fellowship
Pays : International
Informations de copyright
© 2019, Shanmuganathan et al.
Déclaration de conflit d'intérêts
VS, NS, AM, JC, KB, FC, OB, BB, KK, Gv, RB No competing interests declared
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