Evidence that regulation of intramembrane proteolysis is mediated by substrate gating during sporulation in Bacillus subtilis

Ramírez-Guadiana, FH; Rodrigues, CDA; Marquis, KA; Campo, N; Barajas-Ornelas, RDC; Brock, K; Marks, DS; Kruse, AC; Rudner, DZ

Evidence that regulation of intramembrane proteolysis is mediated by substrate gating during sporulation in Bacillus subtilis

Ramírez-Guadiana, FH

Rodrigues, CDA

Marquis, KA Campo, N Barajas-Ornelas, RDC Brock, K Marks, DS Kruse, AC

Rudner, DZ

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Publication Type:: Journal Article
Citation:: PLoS Genetics, 2018, 14 (11)
Issue Date:: 2018-11-01

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Field	Value	Language
dc.contributor.author	Ramírez-Guadiana, FH https://orcid.org/0000-0002-2814-5274	en_US
dc.contributor.author	Rodrigues, CDA https://orcid.org/0000-0003-4541-0145	en_US
dc.contributor.author	Marquis, KA	en_US
dc.contributor.author	Campo, N	en_US
dc.contributor.author	Barajas-Ornelas, RDC	en_US
dc.contributor.author	Brock, K	en_US
dc.contributor.author	Marks, DS	en_US
dc.contributor.author	Kruse, AC https://orcid.org/0000-0002-1467-1222	en_US
dc.contributor.author	Rudner, DZ https://orcid.org/0000-0002-0236-7143	en_US
dc.date.available	2018-10-10	en_US
dc.date.issued	2018-11-01	en_US
dc.identifier.citation	PLoS Genetics, 2018, 14 (11)	en_US
dc.identifier.issn	1553-7390	en_US
dc.identifier.uri	http://hdl.handle.net/10453/129336
dc.description.abstract	© 2018 Ramírez-Guadiana et al. http://creativecommons.org/licenses/by/4.0/. During the morphological process of sporulation in Bacillus subtilis two adjacent daughter cells (called the mother cell and forespore) follow different programs of gene expression that are linked to each other by signal transduction pathways. At a late stage in development, a signaling pathway emanating from the forespore triggers the proteolytic activation of the mother cell transcription factor σK. Cleavage of pro-σK to its mature and active form is catalyzed by the intramembrane cleaving metalloprotease SpoIVFB (B), a Site-2 Protease (S2P) family member. B is held inactive by two mother-cell membrane proteins SpoIVFA (A) and BofA. Activation of pro-σK processing requires a site-1 signaling protease SpoIVB (IVB) that is secreted from the forespore into the space between the two cells. IVB cleaves the extracellular domain of A but how this cleavage activates intramembrane proteolysis has remained unclear. Structural studies of the Methanocaldococcus jannaschii S2P homolog identified closed (substrate-occluded) and open (substrate-accessible) conformations of the protease, but the biological relevance of these conformations has not been established. Here, using co-immunoprecipitation and fluorescence microscopy, we show that stable association between the membrane-embedded protease and its substrate requires IVB signaling. We further show that the cytoplasmic cystathionine-β-synthase (CBS) domain of the B protease is not critical for this interaction or for pro-σK processing, suggesting the IVB-dependent interaction site is in the membrane protease domain. Finally, we provide evidence that the B protease domain adopts both open and closed conformations in vivo. Collectively, our data support a substrate-gating model in which IVB-dependent cleavage of A on one side of the membrane triggers a conformational change in the membrane-embedded protease from a closed to an open state allowing pro-σK access to the caged interior of the protease.	en_US
dc.relation.ispartof	PLoS Genetics	en_US
dc.relation.isbasedon	10.1371/journal.pgen.1007753	en_US
dc.subject.classification	Developmental Biology	en_US
dc.subject.mesh	Spores	en_US
dc.subject.mesh	Bacillus subtilis	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Membrane Proteins	en_US
dc.subject.mesh	Gene Expression	en_US
dc.subject.mesh	Protein Conformation	en_US
dc.subject.mesh	Protein Transport	en_US
dc.subject.mesh	Models, Molecular	en_US
dc.subject.mesh	Protein Stability	en_US
dc.subject.mesh	Proteolysis	en_US
dc.title	Evidence that regulation of intramembrane proteolysis is mediated by substrate gating during sporulation in Bacillus subtilis	en_US
dc.type	Journal Article
utslib.citation.volume	11	en_US
utslib.citation.volume	14	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	0604 Genetics	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.issue	11	en_US
pubs.publication-status	Published	en_US
pubs.volume	14	en_US

Abstract:

© 2018 Ramírez-Guadiana et al. http://creativecommons.org/licenses/by/4.0/. During the morphological process of sporulation in Bacillus subtilis two adjacent daughter cells (called the mother cell and forespore) follow different programs of gene expression that are linked to each other by signal transduction pathways. At a late stage in development, a signaling pathway emanating from the forespore triggers the proteolytic activation of the mother cell transcription factor σK. Cleavage of pro-σK to its mature and active form is catalyzed by the intramembrane cleaving metalloprotease SpoIVFB (B), a Site-2 Protease (S2P) family member. B is held inactive by two mother-cell membrane proteins SpoIVFA (A) and BofA. Activation of pro-σK processing requires a site-1 signaling protease SpoIVB (IVB) that is secreted from the forespore into the space between the two cells. IVB cleaves the extracellular domain of A but how this cleavage activates intramembrane proteolysis has remained unclear. Structural studies of the Methanocaldococcus jannaschii S2P homolog identified closed (substrate-occluded) and open (substrate-accessible) conformations of the protease, but the biological relevance of these conformations has not been established. Here, using co-immunoprecipitation and fluorescence microscopy, we show that stable association between the membrane-embedded protease and its substrate requires IVB signaling. We further show that the cytoplasmic cystathionine-β-synthase (CBS) domain of the B protease is not critical for this interaction or for pro-σK processing, suggesting the IVB-dependent interaction site is in the membrane protease domain. Finally, we provide evidence that the B protease domain adopts both open and closed conformations in vivo. Collectively, our data support a substrate-gating model in which IVB-dependent cleavage of A on one side of the membrane triggers a conformational change in the membrane-embedded protease from a closed to an open state allowing pro-σK access to the caged interior of the protease.

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http://hdl.handle.net/10453/129336