Multiple GTPases participate in the assembly of the large ribosomal subunit in Bacillus subtilis.
GTPases have been demonstrated to be necessary for the proper assembly of the ribosome in bacteria and eukaryotes. Here, we show that the essential GTPases YphC and YsxC are required for large ribosomal subunit biogenesis in Bacillus subtilis. Sucrose density gradient centrifugation of large ribosomal subunits isolated from YphC-depleted cells and YsxC-depleted cells indicates that they are similar to the 45S intermediate previously identified in RbgA-depleted cells. The sedimentation of the large-subunit intermediate isolated from YphC-depleted cells was identical to the intermediate found in RbgA-depleted cells, while the intermediate isolated from YsxC-depleted cells sedimented slightly slower than 45S, suggesting that it is a novel intermediate. Analysis of the protein composition of the large-subunit intermediates isolated from either YphC-depleted cells or YsxC-depleted cells indicated that L16 and L36 are missing. Purified YphC and YsxC are able to interact with the ribosome in vitro, supporting a direct role for these two proteins in the assembly of the 50S subunit. Our results indicate that, as has been demonstrated for Saccharomyces cerevisiae ribosome biogenesis, bacterial 50S ribosome assembly requires the function of multiple essential GTPases.
Schaefer
Laura
L
Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan 48824, USA.
Uicker
William C
WC
Wicker-Planquart
Catherine
C
Foucher
Anne-Emmanuelle
AE
Jault
Jean-Michel
JM
Britton
Robert A
RA
D001412:Bacillus subtilis / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000502:physiology*; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D017353:Gene Deletion; D005798:Genes, Bacterial; D008958:Models, Molecular; D012269:Ribosomal Proteins / Q000096:biosynthesis*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*
D001426:Bacterial Proteins; D012269:Ribosomal Proteins; D020558:GTP Phosphohydrolases