Essential autoproteolysis of bacterial anti-σ factor RsgI for transmembrane signal transduction

Chen, Chao; Dong, Sheng; Yu, Zhaoli; Qiao, Yichen; Li, Jie; Ding, Xiaoke; Li, Renmin; Lin, Jinzhong; Bayer, Edward A.; Liu, Ya-Jun; Cui, Qiu; Feng, Yingang

doi:10.1126/sciadv.adg4846

Cited by 12 publications

(4 citation statements)

References 65 publications

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“…While this study was under review, Chen et al reported NMR and crystal structures of the SEAL domains from H. thermocellum RsgI1, RsgI2, and RsgI6 and the finding that these domains undergo autoproteolysis, as we have described here ( 30 ). The structures of the cleaved H. thermocellum SEAL domains have a similar fold to B. subtilis RsgI GGG and contain analogous extensions of the β-strands proximal to the cleavage site.…”

Section: Discussionsupporting

confidence: 57%

See 1 more Smart Citation

Bacterial SEAL domains undergo autoproteolysis and function in regulated intramembrane proteolysis

Brogan,

Habib,

Hobbs

et al. 2023

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

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Gram-positive bacteria use SigI/RsgI-family sigma factor/anti-sigma factor pairs to sense and respond to cell wall defects and plant polysaccharides. In Bacillus subtilis, this signal transduction pathway involves regulated intramembrane proteolysis (RIP) of the membrane-anchored anti-sigma factor RsgI. However, unlike most RIP signaling pathways, site-1 cleavage of RsgI on the extracytoplasmic side of the membrane is constitutive and the cleavage products remain stably associated, preventing intramembrane proteolysis. The regulated step in this pathway is their dissociation, which is hypothesized to involve mechanical force. Release of the ectodomain enables intramembrane cleavage by the RasP site-2 protease and activation of SigI. The constitutive site-1 protease has not been identified for any RsgI homolog. Here, we report that RsgI’s extracytoplasmic domain has structural and functional similarities to eukaryotic SEA domains that undergo autoproteolysis and have been implicated in mechanotransduction. We show that site-1 proteolysis in B. subtilis and Clostridial RsgI family members is mediated by enzyme-independent autoproteolysis of these SEA-like domains. Importantly, the site of proteolysis enables retention of the ectodomain through an undisrupted β-sheet that spans the two cleavage products. Autoproteolysis can be abrogated by relief of conformational strain in the scissile loop, in a mechanism analogous to eukaryotic SEA domains. Collectively, our data support the model that RsgI–SigI signaling is mediated by mechanotransduction in a manner that has striking parallels with eukaryotic mechanotransducive signaling pathways.

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Section: Discussionsupporting

confidence: 57%

“…3 A ). A recent report provides strong support for this model and implicates the H. thermocellum RasP homolog as the site-2 protease ( 30 ).…”

Section: Resultsmentioning

confidence: 88%

Bacterial SEAL domains undergo autoproteolysis and function in regulated intramembrane proteolysis

Brogan,

Habib,

Hobbs

et al. 2023

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

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“…NMR structure calculation procedures were similar to those previously described (Chen et al, 2023; Wei et al, 2019). Initial dDoc_0689 structures were calculated using the CYANA (Herrmann et al, 2002) software with the NOE peak lists from the 3D 15 N‐edited and 13 C‐edited NOESY HSQC spectra and dihedral angle restraints derived from the chemical shifts using the program TALOS‐N (Shen & Bax, 2015).…”

Section: Methodsmentioning

confidence: 99%

A cellulosomal double‐dockerin module from Clostridium thermocellum shows distinct structural and cohesin‐binding features

Chen,

Yang,

Dong

et al. 2024

Protein Science

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Cellulosomes are intricate cellulose‐degrading multi‐enzymatic complexes produced by anaerobic bacteria, which are valuable for bioenergy development and biotechnology. Cellulosome assembly relies on the selective interaction between cohesin modules in structural scaffolding proteins (scaffoldins) and dockerin modules in enzymes. Although the number of tandem cohesins in the scaffoldins is believed to determine the complexity of the cellulosomes, tandem dockerins also exist, albeit very rare, in some cellulosomal components whose assembly and functional roles are currently unclear. In this study, we characterized the structure and mode of assembly of a tandem bimodular double‐dockerin, which is connected to a putative S8 protease in the cellulosome‐producing bacterium, Clostridium thermocellum. Crystal and NMR structures of the double‐dockerin revealed two typical type I dockerin folds with significant interactions between them. Interaction analysis by isothermal titration calorimetry and NMR titration experiments revealed that the double‐dockerin displays a preference for binding to the cell‐wall anchoring scaffoldin ScaD through the first dockerin with a canonical dual‐binding mode, while the second dockerin module was unable to bind to any of the tested cohesins. Surprisingly, the double‐dockerin showed a much higher affinity to a cohesin from the CipC scaffoldin of Clostridium cellulolyticum than to the resident cohesins from C. thermocellum. These results contribute valuable insights into the structure and assembly of the double‐dockerin module, and provide the basis for further functional studies on multiple‐dockerin modules and cellulosomal proteases, thus highlighting the complexity and diversity of cellulosomal components.

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“…The modules within the cellulosome are connected by flexible linkers, allowing the cellulosome to undergo conformational changes according to the substrate, thus degrading the substrate better [162]. Additionally, bacteria express genes of cellulosomal components dynamically based on the type of substrate and adapt to the different substrate compositions [162][163][164][165][166]. Compared to free cellulases, cellulosomes not only exhibit a stronger tolerance to chemical inhibitors such as formate, lactate, and furfural present in the hydrolysate but also show a higher ethanol tolerance and thermostability [157].…”

Section: Saccharification Processmentioning

confidence: 99%

Composition of Lignocellulose Hydrolysate in Different Biorefinery Strategies: Nutrients and Inhibitors

Wang,

Zhang,

Cui

et al. 2024

Molecules

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The hydrolysis and biotransformation of lignocellulose, i.e., biorefinery, can provide human beings with biofuels, bio-based chemicals, and materials, and is an important technology to solve the fossil energy crisis and promote global sustainable development. Biorefinery involves steps such as pretreatment, saccharification, and fermentation, and researchers have developed a variety of biorefinery strategies to optimize the process and reduce process costs in recent years. Lignocellulosic hydrolysates are platforms that connect the saccharification process and downstream fermentation. The hydrolysate composition is closely related to biomass raw materials, the pretreatment process, and the choice of biorefining strategies, and provides not only nutrients but also possible inhibitors for downstream fermentation. In this review, we summarized the effects of each stage of lignocellulosic biorefinery on nutrients and possible inhibitors, analyzed the huge differences in nutrient retention and inhibitor generation among various biorefinery strategies, and emphasized that all steps in lignocellulose biorefinery need to be considered comprehensively to achieve maximum nutrient retention and optimal control of inhibitors at low cost, to provide a reference for the development of biomass energy and chemicals.

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Essential autoproteolysis of bacterial anti-σ factor RsgI for transmembrane signal transduction

Cited by 12 publications

References 65 publications

Bacterial SEAL domains undergo autoproteolysis and function in regulated intramembrane proteolysis

Bacterial SEAL domains undergo autoproteolysis and function in regulated intramembrane proteolysis

A cellulosomal double‐dockerin module from Clostridium thermocellum shows distinct structural and cohesin‐binding features

Composition of Lignocellulose Hydrolysate in Different Biorefinery Strategies: Nutrients and Inhibitors

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