The DEAD-box RNA helicase SrmB is involved in the assembly of 50S ribosomal subunits in Escherichia coli.
Ribosome assembly in Escherichia coli involves 54 ribosomal proteins and three RNAs. Whereas functional subunits can be reconstituted in vitro from the isolated components, this process requires long incubation times and high temperatures compared with the in vivo situation, suggesting that non-ribosomal factors facilitate assembly in vivo. Here, we show that SrmB, a putative DEAD-box RNA helicase, is involved in ribosome assembly. The deletion of the srmB gene causes a slow-growth phenotype at low temperature. Polysome profile analyses of the corresponding cells reveal a deficit in free 50S ribosomal subunits and the accumulation of a new particle sedimenting around 40S. Analysis of the ribosomal RNA and protein contents of the 40S particle indicates that it represents a large subunit that is incompletely assembled. In particular, it lacks L13, one of the five ribosomal proteins that are essential for the early assembly step in vitro. Sucrose gradient fractionation also shows that, in wild-type cells, SrmB associates with a pre50S particle. From our results, we propose that SrmB is involved in an early step of 50S assembly that is necessary for the binding of L13. This step may consist of a structural rearrangement that, at low temperature, cannot occur without the assistance of this putative RNA helicase.
Charollais
Julie
J
Laboratoire de Génétique Moléculaire, CNRS UMR8541, Ecole Normale Supérieure, 46 rue d'Ulm, 75230 Paris Cedex 05, France.
Pflieger
Delphine
D
Vinh
Joëlle
J
Dreyfus
Marc
M
Iost
Isabelle
I
D002499:Centrifugation, Density Gradient; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D017353:Gene Deletion; D013058:Mass Spectrometry; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D020365:RNA Helicases / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; D020365:RNA Helicases