TY - JOUR
T1 - Conserved heterodimeric GTPase Rbg1/Tma46 promotes efficient translation in eukaryotic cells
AU - Zeng, Fuxing
AU - Li, Xin
AU - Pires-Alves, Melissa
AU - Chen, Xin
AU - Hawk, Christopher W.
AU - Jin, Hong
N1 - Funding Information:
We thank Prof. Peter Moore at Yale University for critical comments on the manuscript. We also thank the Cryo-EM Center at Southern University of Science and Technology, Junyi Jiang and Qingrong Li in the F. Zeng laboratory, Dr. Bridget Carragher and Dr. Ed Eng at the National Center for CryoEM Access and Training (NCCAT) and Dr. Valerie Tokars at Northwestern University for their help on cryoEM data collection, the core Research Facility at Southern University of Science and Technology for flow cytometry, Roy J. Carver Biotechnology Center at the University of Illinois at Urbana-Champaign for sequencing, and members in the Jin laboratory for helpful discussions. H.J. acknowledges support from the National Institute of General Medical Sciences of the NIH (R01-GM120552). H.J. and F.Z. designed the study. F.Z. X.L. M.P.-A. and C.W.H. performed the experiments. F.Z. analyzed the data, and X.C. helped on the 5P-Seq data analysis. F.Z. and C.W.H. helped with manuscript preparation. H.J. wrote the paper. All authors discussed the final manuscript. The authors declare no competing interests, One or more of the authors of this paper self-identifies as an underrepresented ethnic minority in science. One or more of the authors of this paper self-identifies as living with a disability.
Funding Information:
We thank Prof. Peter Moore at Yale University for critical comments on the manuscript. We also thank the Cryo-EM Center at Southern University of Science and Technology, Junyi Jiang and Qingrong Li in the F. Zeng laboratory, Dr. Bridget Carragher and Dr. Ed Eng at the National Center for CryoEM Access and Training (NCCAT) and Dr. Valerie Tokars at Northwestern University for their help on cryoEM data collection, the core Research Facility at Southern University of Science and Technology for flow cytometry, Roy J. Carver Biotechnology Center at the University of Illinois at Urbana-Champaign for sequencing, and members in the Jin laboratory for helpful discussions. H.J. acknowledges support from the National Institute of General Medical Sciences of the NIH ( R01-GM120552 ).
Publisher Copyright:
© 2021 The Authors
PY - 2021/10/26
Y1 - 2021/10/26
N2 - Conserved developmentally regulated guanosine triphosphate (GTP)-binding proteins (Drgs) and their binding partner Drg family regulatory proteins (Dfrps) are important for embryonic development, cellular growth control, differentiation, and proliferation. Here, we report that the yeast Drg1/Dfrp1 ortholog Rbg1/Tma46 facilitates translational initiation, elongation, and termination by suppressing prolonged ribosome pausing. Consistent with the genome-wide observations, deletion of Rbg1 exacerbates the growth defect resulting from translation stalling, and Rbg1 stabilizes mRNAs against no-go decay. Furthermore, we provide a cryoelectron microscopy (cryo-EM) structure of the 80S ribosome bound with Rbg1/Tma46 that reveals the molecular interactions responsible for Rbg1/Tma46 function. The Rbg1 subunit binds to the GTPase association center of the ribosome and the A-tRNA, and the N-terminal zinc finger domain of the Tma46 subunit binds to the 40S, establishing an interaction critical for the ribosomal association. Our results answer the fundamental question of how a paused ribosome resumes translation and show that Drg1/Dfrp1 play a critical role in ensuring orderly translation.
AB - Conserved developmentally regulated guanosine triphosphate (GTP)-binding proteins (Drgs) and their binding partner Drg family regulatory proteins (Dfrps) are important for embryonic development, cellular growth control, differentiation, and proliferation. Here, we report that the yeast Drg1/Dfrp1 ortholog Rbg1/Tma46 facilitates translational initiation, elongation, and termination by suppressing prolonged ribosome pausing. Consistent with the genome-wide observations, deletion of Rbg1 exacerbates the growth defect resulting from translation stalling, and Rbg1 stabilizes mRNAs against no-go decay. Furthermore, we provide a cryoelectron microscopy (cryo-EM) structure of the 80S ribosome bound with Rbg1/Tma46 that reveals the molecular interactions responsible for Rbg1/Tma46 function. The Rbg1 subunit binds to the GTPase association center of the ribosome and the A-tRNA, and the N-terminal zinc finger domain of the Tma46 subunit binds to the 40S, establishing an interaction critical for the ribosomal association. Our results answer the fundamental question of how a paused ribosome resumes translation and show that Drg1/Dfrp1 play a critical role in ensuring orderly translation.
KW - Dfrp
KW - Drg
KW - Drg-family regulatory proteins
KW - developmentally-regulated GTP-binding proteins
KW - protein homeostasis
KW - ribosome
KW - ribosome stalling
KW - translation
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UR - http://www.scopus.com/inward/citedby.url?scp=85117904797&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2021.109877
DO - 10.1016/j.celrep.2021.109877
M3 - Article
C2 - 34706231
AN - SCOPUS:85117904797
SN - 2211-1247
VL - 37
JO - Cell Reports
JF - Cell Reports
IS - 4
M1 - 109877
ER -