Suppressor mutations in ribosomal proteins and FliY restore Bacillus subtilis swarming motility in the absence of EF-P

Hummels, Katherine R.; Kearns, Daniel B.

doi:10.1371/journal.pgen.1008179

Cited by 20 publications

(14 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To determine how CwlQ might promote swarming, we sought to isolate spontaneous suppressors that restored motility to a cwlQ mutant upon prolonged incubation on a swarm agar plate. Unlike regulatory mutants defective in swarming (14,41,50,54,55), no spontaneous swarming-proficient suppressor ever emerged as a flare from the non-motile colony of the cwlQ mutant, even after 48 hours of incubation. To directly test the hypothesis that flagellar number was limiting in the cwlQ mutant, a double mutant was generated that was simultaneously defective in cwlQ and smiA encoding SmiA, a specific adaptor protein for the regulatory proteolysis of SwrA (46).…”

Section: Resultsmentioning

confidence: 99%

CwlQ Is Required for Swarming Motility but Not Flagellar Assembly in Bacillus subtilis

2021

Self Cite

View full text Add to dashboard Cite

Lytic enzymes play an essential role in the remodeling of bacterial peptidoglycan (PG), an extracellular mesh-like structure that retains the membrane in the context of high internal osmotic pressure. Peptidoglycan (PG) must be unfailingly stable to preserve cell integrity but must also be dynamically remodeled for the cell grow, divide and insert macromolecular machines. The flagellum is one such macromolecular machine that transits the PG and flagellar insertion is aided by localized activity of a dedicated PG lyase in Gram-negative bacteria. To date, there is no known dedicated lyase in Gram-positive bacteria for the insertion of flagella and here we take a reverse-genetic candidate-gene approach and find that cells mutated for the lytic transglycosylase CwlQ exhibited a severe defect in flagellar-dependent swarming motility. We further show that CwlQ was expressed by the motility sigma factor SigD and was secreted by the type III secretion system housed inside the flagellum. Nonetheless, cells mutated for CwlQ remained proficient for flagellar biosynthesis even when mutated in combination with four other lyases related to motility (LytC, LytD, LytF, and CwlO). The PG lyase or lyases essential for flagellar synthesis in B. subtilis, if any, remains unknown. IMPORTANCE Bacteria are surrounded by a wall of peptidoglycan and early work in Bacillus subtilis was the first to suggest that bacteria needed to enzymatically remodel the wall to permit insertion of the flagellum. No PG remodeling enzyme alone or in combination however, has been found to be essential for flagellar assembly in B. subtilis. Here we take a reverse genetic candidate gene approach and find that the PG lytic transglycosylase CwlQ is required for swarming motility. Subsequent characterization determined that while CwlQ was co-expressed with motility genes and is secreted by the flagellar secretion apparatus, it was not required for flagellar synthesis. The PG lyase needed for flagellar assembly in B. subtilis remains unknown.

show abstract

Section: Resultsmentioning

confidence: 99%

CwlQ Is Required for Swarming Motility but Not Flagellar Assembly in Bacillus subtilis

2021

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

CwlQ is required for swarming motility but not flagellar assembly inBacillus subtilis

Kearns

Sánchez

Dunn

2021

Preprint

Self Cite

View full text Add to dashboard Cite

Hydrolytic enzymes play an essential role in the remodeling of bacterial peptidoglycan (PG), an extracellular mesh-like structure that retains the membrane in the context of high internal osmotic pressure. Peptidoglycan (PG) integrity must be unfailingly stable to preserve cell integrity but must also be dynamically remodeled for the cell grow, divide and insert macromolecular machines. The flagellum is one such macromolecular machine that transits the PG and the insertion of which is aided by localized activity of a dedicated PG hydrolase in Gram-negative bacteria. To date, there is no known dedicated hydrolase in Gram-positive bacteria for insertion of flagella and here we take a reverse-genetic candidate-gene approach to find that cells mutated for the lytic transglycosylase CwlQ exhibited a severe defect in flagellar dependent swarming motility. We show that CwlQ required its active site to promote swarming, was expressed by the motility sigma factor SigD, and was secreted by the type III secretion system housed inside the flagellum. Nonetheless, cells mutated for CwlQ remained proficient for flagellar biosynthesis even when mutated in combination with four other hydrolases related to motility (LytC, LytD, LytF, and CwlO). The PG hydrolase essential for flagellar synthesis in B. subtilis, if any, remains unknown.

show abstract

“…Research involving EFP and di-prolyl motifs has largely focused on individual species [24,25,27,29] or proteins [67]. In contrast, we surveyed these motifs in a broad range of genotypically and phenotypically diverse bacteria.…”

Section: Discussionmentioning

confidence: 99%

“…The importance of EFP and its mitigation of ribosome stalling is underscored by the strong phenotypes caused by its loss. These include diminished growth rate [20][21][22][23][24] , loss of motility [25], loss of virulence [20,22,24], reduced antibiotic resistance [24,26], and in some cases, cell death [27].…”

Section: Introductionmentioning

confidence: 99%

Translation stalling proline motifs are enriched in slow-growing, thermophilic, and multicellular bacteria

Brewer

Wagner

2021

Preprint

View full text Add to dashboard Cite

Rapid bacterial growth depends on the speed at which ribosomes can translate mRNA into proteins. mRNAs that encode successive stretches of proline can cause ribosomes to stall, substantially reducing translation speed. Such stalling is especially detrimental for species that must grow and divide rapidly. Here we focus on di-prolyl motifs (XXPPX) and ask whether their incidence varies with growth rate. To find out we conducted a broad survey of such motifs in >3000 bacterial genomes across 36 phyla. Indeed, fast-growing species encode fewer motifs than slow-growing species, especially in highly expressed proteins. We also found many di-prolyl motifs within thermophiles, where prolines can help maintain proteome stability. Moreover, bacteria with complex, multicellular lifecycles also encode many di-prolyl motifs. This is especially evident in the slow-growing phylum Myxococcota. Bacteria in this phylum encode many serine-threonine kinases, and many di-prolyl motifs at potential phosphorylation sites within these kinases. Serine-threonine kinases are involved in cell signaling and help regulate developmental processes linked to multicellularity in the Myxococcota. Altogether, our observations suggest that weakened selection on translational rate, whether due to slow or thermophilic growth, may allow di-prolyl motifs to take on new roles in biological processes that are unrelated to translational rate.

show abstract

Suppressor mutations in ribosomal proteins and FliY restore Bacillus subtilis swarming motility in the absence of EF-P

Cited by 20 publications

References 67 publications

CwlQ Is Required for Swarming Motility but Not Flagellar Assembly in Bacillus subtilis

CwlQ Is Required for Swarming Motility but Not Flagellar Assembly in Bacillus subtilis

CwlQ is required for swarming motility but not flagellar assembly inBacillus subtilis

Translation stalling proline motifs are enriched in slow-growing, thermophilic, and multicellular bacteria

Contact Info

Product

Resources

About