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Nucleic Acids Res. 2016 Sep 30;44(17):8442-55. doi: 10.1093/nar/gkw678. Epub 2016 Aug 2.

YphC and YsxC GTPases assist the maturation of the central protuberance, GTPase associated region and functional core of the 50S ribosomal subunit.

Author information

1
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario L8S4K1, Canada M.G. DeGroote Institute for Infectious Diseases Research, McMaster University, Hamilton, Ontario L8S4K1, Canada.
2
Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA Department of Chemistry and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
3
Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX 77030, USA Center for Metagenomics and Microbiome Research, Baylor College of Medicine, Houston, TX 77030, USA.
4
Molecular Structure and Function Program, The Hospital for Sick Children,Toronto, Ontario M5G 0A4, Canada.
5
Molecular Structure and Function Program, The Hospital for Sick Children,Toronto, Ontario M5G 0A4, Canada Department of Biochemistry, University of Toronto, Toronto, Ontario M5S 1A8, Canada Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 1L7, Canada.
6
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario L8S4K1, Canada M.G. DeGroote Institute for Infectious Diseases Research, McMaster University, Hamilton, Ontario L8S4K1, Canada ortegaj@mcmaster.ca.

Abstract

YphC and YsxC are GTPases in Bacillus subtilis that facilitate the assembly of the 50S ribosomal subunit, however their roles in this process are still uncharacterized. To explore their function, we used strains in which the only copy of the yphC or ysxC genes were under the control of an inducible promoter. Under depletion conditions, they accumulated incomplete ribosomal subunits that we named 45SYphC and 44.5SYsxC particles. Quantitative mass spectrometry analysis and the 5-6 Å resolution cryo-EM maps of the 45SYphC and 44.5SYsxC particles revealed that the two GTPases participate in the maturation of the central protuberance, GTPase associated region and key RNA helices in the A, P and E functional sites of the 50S subunit. We observed that YphC and YsxC bind specifically to the two immature particles, suggesting that they represent either on-pathway intermediates or that their structure has not significantly diverged from that of the actual substrate. These results describe the nature of these immature particles, a widely used tool to study the assembly process of the ribosome. They also provide the first insights into the function of YphC and YsxC in 50S subunit assembly and are consistent with this process occurring through multiple parallel pathways, as it has been described for the 30S subunit.

PMID:
27484475
PMCID:
PMC5041480
DOI:
10.1093/nar/gkw678
[Indexed for MEDLINE]
Free PMC Article

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