title: SrmB, a DEAD-box helicase involved in Escherichia coli ribosome assembly, is specifically targeted to 23S rRNA in vivo.
abstract: DEAD-box proteins play specific roles in remodeling RNA or ribonucleoprotein complexes. Yet, in vitro, they generally behave as nonspecific RNA-dependent ATPases, raising the question of what determines their specificity in vivo. SrmB, one of the five Escherichia coli DEAD-box proteins, participates in the assembly of the large ribosomal subunit. Moreover, when overexpressed, it compensates for a mutation in L24, the ribosomal protein (r-protein) thought to initiate assembly. Here, using the tandem affinity purification (TAP) procedure, we show that SrmB forms a complex with r-proteins L4, L24 and a region near the 5'-end of 23S rRNA that binds these proteins. In vitro reconstitution experiments show that the stability of this complex reflects cooperative interactions of SrmB with L4, L24 and rRNA. These observations are consistent with an early role of SrmB in assembly and explain the genetic link between SrmB and L24. Besides its catalytic core, SrmB possesses a nonconserved C-terminal extension that, we show, is not essential for SrmB function and specificity. In this regard, SrmB differs from DbpA, another DEAD-box protein involved in ribosome assembly.
authors.affiliation: Laboratoire de Génétique Moléculaire, CNRS UMR8541, Ecole Normale Supérieure, 75230 Paris Cedex 05, France.
authors.forename: Dmitrii
authors.initials: D
authors.lastname: Trubetskoy
authors.forename: Florence
authors.initials: F
authors.lastname: Proux
authors.forename: Frédéric
authors.initials: F
authors.lastname: Allemand
authors.forename: Marc
authors.initials: M
authors.lastname: Dreyfus
authors.forename: Isabelle
authors.initials: I
authors.lastname: Iost
mesh_terms: D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D012338:RNA, Ribosomal, 23S / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D012338:RNA, Ribosomal, 23S; D012261:Ribonucleoproteins; C545660:SrmB protein, E coli; D053487:DEAD-box RNA Helicases