Multiple GTPases Participate in the Assembly of the Large Ribosomal Subunit in
            <i>Bacillus subtilis</i>

Schaefer, Laura; Uicker, William C.; Wicker‐Planquart, Catherine; Foucher, Anne-Emmanuelle; Jault, Jean‐Michel; Britton, Robert A.

doi:10.1128/jb.01213-06

Cited by 73 publications

(112 citation statements)

References 35 publications

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“…RbgA/YlqF is an essential GTPase required for the proliferation of B. subtilis (36,37). Depletion of RbgA or expression of a dominant-negative mutant of RbgA resulted in accumulation of the 45S particles (36)(37)(38)(39). However, unlike the E. coli 45S precursor, the B. subtilis 45S particle does not alter its sedimentation coefficient, even at 10 mM Mg 2+ (36,39).…”

Section: Discussionmentioning

confidence: 99%

Single methylation of 23S rRNA triggers late steps of 50S ribosomal subunit assembly

Arai

Ishiguro

Kimura

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Ribosome biogenesis requires multiple assembly factors. In Escherichia coli, deletion of RlmE, the methyltransferase responsible for the 2′-O-methyluridine modification at position 2552 (Um2552) in helix 92 of the 23S rRNA, results in slow growth and accumulation of the 45S particle. We demonstrate that the 45S particle that accumulates in ΔrlmE is a genuine precursor that can be assembled into the 50S subunit. Indeed, 50S formation from the 45S precursor could be promoted by RlmE-mediated Um2552 formation in vitro. Ribosomal protein L36 (encoded by rpmJ) was completely absent from the 45S precursor in ΔrlmE, and we observed a strong genetic interaction between rlmE and rpmJ. Structural probing of 23S rRNA and high-salt stripping of 45S components revealed that RlmE-mediated methylation promotes interdomain interactions via the association between helices 92 and 71, stabilized by the single 2′-O-methylation of Um2552, in concert with the incorporation of L36, triggering late steps of 50S subunit assembly.R ibosomes are essential ribonucleoprotein complexes that translate the genetic information encoded by mRNA into protein. Intact bacterial ribosomes, which have a sedimentation coefficient of 70S, consist of large (50S) and small (30S) subunits. Each subunit can be reconstituted in vitro from its individual component ribosomal RNAs and proteins (1-5), indicating that these components contain all of the information necessary to automatically assemble into functional subunits. Because the intermediate particles observed in an in vitro reconstitution of the subunits are similar to those observed in vivo, assembly maps that describe the order of ribosomal protein binding in vitro are useful tools for understanding ribosome biogenesis in the cell (6). However, assembly is slower in vitro than in vivo, and nonphysiological conditions, such as high-salt concentration and high temperature, are required for in vitro assembly.In the cell, ribosome assembly is coordinated with the transcription and processing of large precursor rRNAs encoded by the rrn operon (3, 7). Once transcription starts, nascent transcripts rapidly form local secondary and tertiary structures that serve as binding sites for the primary ribosomal proteins, thereby initiating ribosome assembly. However, hierarchical assembly starting at the 5′-terminal domains of the 16S and 23S rRNAs is not essential for ribosome formation in the cell (8) because nonribosomal factors, termed "assembly factors," facilitate rapid and efficient assembly of ribosomal particles. Ribosome biogenesis is an energy-consuming process in which a number of assembly factors, including RNA helicases, GTPases, protein chaperones, and rRNA-modifying enzymes, support efficient assembly of each subunit (6, 9, 10). Individual mutation or deletion of several assembly factors causes accumulation of immature 50S subunits that sediment at 40S or 45S. For example, the RNA helicase SrmB participates in the early stage of 50S assembly (11,12); deletion of srmB results in accumulation of a 40S ...

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Section: Discussionmentioning

confidence: 99%

Single methylation of 23S rRNA triggers late steps of 50S ribosomal subunit assembly

Arai

Ishiguro

Kimura

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…The intermediate observed upon YihA depletion has a slightly slower sedimentation rate than the 45S intermediates observed when RbgA and EngA are depleted. [46][47][48] In contrast, depletion of Obg leads to accumulation of a 50S pre-ribosome that is unstable at low Mg 21 concentrations. 49 The increasing sedimentation rate observed with intermediates accumulating when YihA, RbgA, EngA, and Obg are depleted, suggest a preliminary order to the action of these GTPases with YihA acting before RbgA and EngA, and Obg being the last GTPase to act on assembling 50S subunits ( Figure 4B).…”

Section: Gtpases Constitute a Large And Diverse Class Of Ribosome Assmentioning

confidence: 99%

“…In addition to these GTPases, bacteria also encode additional GTPases involved in 50S assembly: YihA and EngA (Der). 46,50,51 In bacteria, depletion of these GTPases leads to the accumulation of distinct intermediates, which are stable in sucrose gradients and can be purified and analyzed for their protein content. The intermediate observed upon YihA depletion has a slightly slower sedimentation rate than the 45S intermediates observed when RbgA and EngA are depleted.…”

Section: Gtpases Constitute a Large And Diverse Class Of Ribosome Assmentioning

confidence: 99%

Role of GTPases in ribosome assembly

Karbstein

2007

Biopolymers

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GTPases are a universally conserved class of regulatory proteins involved in such diverse cellular functions as signal transduction, translation, cytoskeleton formation, and intracellular transport. GTPases are also required for ribosome assembly in eukaryotes and bacteria, where they present themselves as possible regulatory molecules. Strikingly, in bacteria they represent the largest class of essential assembly factors. A review of their common structural, biochemical and genetic interactions is presented and integrated with models for their function in ribosome assembly. © 2007 Wiley Periodicals, Inc. Biopolymers 87: 1–11, 2007This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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“…YsxC interacts in vitro with L1, L6 and L7/12 [6], as well as with rRNA [7]. Depletion of YsxC in B. subtilis leads to accumulation of immature ribosomal subunit [8], suggesting that YsxC is required for 50S ribosomal subunit biogenesis.…”

Section: Introductionmentioning

confidence: 99%

The C‐terminal α‐helix of YsxC is essential for its binding to 50S ribosome and rRNAs

2015

Self Cite

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a b s t r a c tYsxC is an essential P-loop GTPase that interacts with the 50S subunit of the ribosome. The putative implication in ribosome binding of two basic clusters of YsxC, a conserved positively charged patch including R31, R116, H117 and K146 lying adjacent to the nucleotide-binding site, and the C-terminal alpha helix, was investigated. C-terminal truncation variants of YsxC were unable to bind to both ribosome and rRNAs, whereas mutations in the other cluster did not affect YsxC binding. Our results indicate that the basic C-terminal region of YsxC is required for its binding to the 50S ribosomal subunit.

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Multiple GTPases Participate in the Assembly of the Large Ribosomal Subunit in Bacillus subtilis

Cited by 73 publications

References 35 publications

Single methylation of 23S rRNA triggers late steps of 50S ribosomal subunit assembly

Single methylation of 23S rRNA triggers late steps of 50S ribosomal subunit assembly

Role of GTPases in ribosome assembly

The C‐terminal α‐helix of YsxC is essential for its binding to 50S ribosome and rRNAs

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