Structural Requirement in Clostridium perfringens Collagenase mRNA 5′ Leader Sequence for Translational Induction through Small RNA-mRNA Base Pairing

Obana, Nozomu; Nomura, Nobuhiko; Nakamura, Kouji

doi:10.1128/jb.00148-13

Cited by 20 publications

(15 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The colA mRNA 5=UTR forms a long stem-loop structure that masks the RBS, thereby inhibiting translation and rendering the transcripts unstable (28). There are two ways to partially melt the secondary structure of the colA mRNA and induce processing in the 5=UTR: (i) base pairing of the sRNA VR-RNA or (ii) elevating temperatures (29). The processed transcripts are stable and translatable because the RBS is free from the secondary structure.…”

Section: Discussionmentioning

confidence: 99%

Role of RNase Y in Clostridium perfringens mRNA Decay and Processing

2017

Self Cite

View full text Add to dashboard Cite

RNase Y is a major endoribonuclease that plays a crucial role in mRNA degradation and processing. We study the role of RNase Y in the Gram-positive anaerobic pathogen Clostridium perfringens, which until now has not been well understood. Our study implies an important role for RNase Y-mediated RNA degradation and processing in virulence gene expression and the physiological development of the organism. We began by constructing an RNase Y conditional knockdown strain in order to observe the importance of RNase Y on growth and virulence. Our resulting transcriptome analysis shows that RNase Y affects the expression of many genes, including toxin-producing genes. We provide data to show that RNase Y depletion repressed several toxin genes in C. perfringens and involved the virR-virS twocomponent system. We also observe evidence that RNase Y is indispensable for processing and stabilizing the transcripts of colA (encoding a major toxin collagenase) and pilA2 (encoding a major pilin component of the type IV pili). Posttranscriptional regulation of colA is known to be mediated by cleavage in the 5= untranslated region (5=UTR), and we observe that RNase Y depletion diminishes colA 5=UTR processing. We show that RNase Y is also involved in the posttranscriptional stabilization of pilA2 mRNA, which is thought to be important for host cell adherence and biofilm formation.IMPORTANCE RNases have important roles in RNA degradation and turnover in all organisms. C. perfringens is a Gram-positive anaerobic spore-forming bacterial pathogen that produces numerous extracellular enzymes and toxins, and it is linked to digestive disorders and disease. A highly conserved endoribonuclease, RNase Y, affects the expression of hundreds of genes, including toxin genes, and studying these effects is useful for understanding C. perfringens specifically and RNases generally. Moreover, RNase Y is involved in processing specific transcripts, and we observed that this processing in C. perfringens results in the stabilization of mRNAs encoding a toxin and bacterial extracellular apparatus pili. Our study shows that RNase activity is associated with gene expression, helping to determine the growth, proliferation, and virulence of C. perfringens.

show abstract

Section: Discussionmentioning

confidence: 99%

Role of RNase Y in Clostridium perfringens mRNA Decay and Processing

2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Total RNA was isolated from C. perfringens and used for Northern blotting as previously described (60). Template DNAs for generation of digoxigenin (DIG)-labeled DNA probes were amplified using the primers listed in Table S2.…”

Section: Methodsmentioning

confidence: 99%

Immunoactive Clostridial Membrane Vesicle Production Is Regulated by a Sporulation Factor

et al. 2017

Self Cite

View full text Add to dashboard Cite

Recently, many Gram-positive bacteria as well as Gram-negative bacteria have been reported to produce membrane vesicles (MVs), but little is known regarding the regulators involved in MV formation. We found that a Gram-positive anaerobic pathogen, Clostridium perfringens, produces MVs predominantly containing membrane proteins and cell wall components. These MVs stimulated proinflammatory cytokine production in mouse macrophage-like cells. We suggested that MVs induced interleukin-6 production through the Toll-like receptor 2 (TLR2) signaling pathway. Thus, the MV could have a role in the bacterium-host interaction and bacterial infection pathogenesis. Moreover, we found that the sporulation master regulator gene spo0A was required for vesiculogenesis. A conserved, phosphorylated aspartate residue of Spo0A was indispensable for MV production, suggesting that the phosphorylation of Spo0A triggers MV production. Multiple orphan sensor kinases necessary for sporulation were also required to maximize MV production. These findings imply that C. perfringens actively produces immunoactive MVs in response to the environment changing, as recognized by membrane-spanning sensor kinases and by modulating the phosphorylation level of Spo0A.

show abstract

“…Total RNA was extracted from C. perfringens derivatives grown at 37°C in GAM broth and used for Northern blot analysis as previously described (16), with digoxigenin (DIG)-labeled DNA probes generated by using DIG-High Prime according to the manufacturer's instructions (17). The template DNA for the toxin gene-specific probes was PCR amplified from strain 13 genomic DNA using the primers described in Abe et al (18).…”

Section: Methodsmentioning

confidence: 99%

A Sporulation Factor Is Involved in the Morphological Change of Clostridium perfringens Biofilms in Response to Temperature

2014

Self Cite

View full text Add to dashboard Cite

Biofilm formation has been associated with bacterial pathogenesis, such as nosocomial and chronic infections, as the resistance of biofilms to environmental stresses has increased. Clostridium perfringens is a Gram-positive spore-forming anaerobic pathogen. This organism survives antibiotic treatment through the formation of biofilms or spores, but the environmental and regulatory factors involved in the biofilm formation remain unclear. Here, we observed that temperature regulates C. perfringens biofilm morphology. At 37°C, C. perfringens adhered to the substrate surface and formed a flat, thin biofilm, herein referred to as adhered biofilm. However, at 25°C, this bacterium did not adhere and produced a threadlike extracellular matrix, forming a viscous, thick biofilm, herein referred to as pellicle biofilm. Pellicle biofilm formation requires the sporulation master regulator, Spo0A, and the toxin regulator, CtrAB, and is enhanced in the absence of the global repressor, AbrB. These transcriptional regulator genes are regulated by each other and temperature. Adhered-biofilm formation requires AbrB and pilA2, which encodes a component of type IV pili (TFP). TFP expression was activated at 37°C and regulated through Spo0A, AbrB, and CtrAB. These results indicate that the morphology of C. perfringens biofilm is dependent on temperature through the differential production of extracellular matrix and the activity of TFP. Moreover, pellicle biofilm formation is involved in sporulation and toxin production. Here, we demonstrated that clostridial biofilm formation is closely associated with sporulation and that the morphological change of the biofilms could play an important role in the pathogenesis of this organism.

show abstract

Structural Requirement in Clostridium perfringens Collagenase mRNA 5′ Leader Sequence for Translational Induction through Small RNA-mRNA Base Pairing

Cited by 20 publications

References 34 publications

Role of RNase Y in Clostridium perfringens mRNA Decay and Processing

Role of RNase Y in Clostridium perfringens mRNA Decay and Processing

Immunoactive Clostridial Membrane Vesicle Production Is Regulated by a Sporulation Factor

A Sporulation Factor Is Involved in the Morphological Change of Clostridium perfringens Biofilms in Response to Temperature

Contact Info

Product

Resources

About