TY - JOUR
T1 - RQT complex dissociates ribosomes collided on endogenous RQC substrate SDD1
AU - Matsuo, Yoshitaka
AU - Tesina, Petr
AU - Nakajima, Shizuka
AU - Mizuno, Masato
AU - Endo, Akinori
AU - Buschauer, Robert
AU - Cheng, Jingdong
AU - Shounai, Okuto
AU - Ikeuchi, Ken
AU - Saeki, Yasushi
AU - Becker, Thomas
AU - Beckmann, Roland
AU - Inada, Toshifumi
N1 - Funding Information:
We thank O. Berninghausen, H. Sieber, C. Ungewickell and S. Rieder for technical support. This work used the Vincent J. Coates Genomics Sequencing Laboratory at UC Berkeley, supported by NIH S10 Instrumentation Grants grant no. OD018174. This study was supported by MEXT/JSPS KAKENHI grant nos. JP18H03977 (T.I.), JP19H05281 (T.I.), JP19K06481 (Y.M.), JP18H03993 (Y.S.) and JP18H05498 (Y.S.); AMED under grant nos. JP19gm1110010 (T.I.) and JP18gm1110003 (Y.S); Research Grants in the Natural Sciences from the Takeda Foundation (T.I.) and by Research Grants in the Medical Sciences from Kato Memorial Bioscience Foundation (Y.M.) and by a grant of the Deutsche Forschungsgemeinschaft (DFG; BE1814/15-1) to R. Beckmann and a PhD fellowship from Boehringer Ingelheim Fonds to R. Buschauer.
PY - 2020/4/1
Y1 - 2020/4/1
N2 - Ribosome-associated quality control (RQC) represents a rescue pathway in eukaryotic cells that is triggered upon translational stalling. Collided ribosomes are recognized for subsequent dissociation followed by degradation of nascent peptides. However, endogenous RQC-inducing sequences and the mechanism underlying the ubiquitin-dependent ribosome dissociation remain poorly understood. Here, we identified SDD1 messenger RNA from Saccharomyces cerevisiae as an endogenous RQC substrate and reveal the mechanism of its mRNA-dependent and nascent peptide−dependent translational stalling. In vitro translation of SDD1 mRNA enabled the reconstitution of Hel2-dependent polyubiquitination of collided disomes and, preferentially, trisomes. The distinct trisome architecture, visualized using cryo-EM, provides the structural basis for the more-efficient recognition by Hel2 compared with that of disomes. Subsequently, the Slh1 helicase subunit of the RQC trigger (RQT) complex preferentially dissociates the first stalled polyubiquitinated ribosome in an ATP-dependent manner. Together, these findings provide fundamental mechanistic insights into RQC and its physiological role in maintaining cellular protein homeostasis.
AB - Ribosome-associated quality control (RQC) represents a rescue pathway in eukaryotic cells that is triggered upon translational stalling. Collided ribosomes are recognized for subsequent dissociation followed by degradation of nascent peptides. However, endogenous RQC-inducing sequences and the mechanism underlying the ubiquitin-dependent ribosome dissociation remain poorly understood. Here, we identified SDD1 messenger RNA from Saccharomyces cerevisiae as an endogenous RQC substrate and reveal the mechanism of its mRNA-dependent and nascent peptide−dependent translational stalling. In vitro translation of SDD1 mRNA enabled the reconstitution of Hel2-dependent polyubiquitination of collided disomes and, preferentially, trisomes. The distinct trisome architecture, visualized using cryo-EM, provides the structural basis for the more-efficient recognition by Hel2 compared with that of disomes. Subsequently, the Slh1 helicase subunit of the RQC trigger (RQT) complex preferentially dissociates the first stalled polyubiquitinated ribosome in an ATP-dependent manner. Together, these findings provide fundamental mechanistic insights into RQC and its physiological role in maintaining cellular protein homeostasis.
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U2 - 10.1038/s41594-020-0393-9
DO - 10.1038/s41594-020-0393-9
M3 - Article
C2 - 32203490
AN - SCOPUS:85082182311
VL - 27
SP - 323
EP - 332
JO - Nature Structural Biology
JF - Nature Structural Biology
SN - 1545-9993
IS - 4
ER -