14 results found
Neef J, Bongiorni C, Schmidt B, et al., 2020, Relative contributions of non-essential Sec pathway components and cell envelope-associated proteases to high-level enzyme secretion by Bacillus subtilis, MICROBIAL CELL FACTORIES, Vol: 19
Walker K, Goosens V, Das A, et al., 2019, Engineered cell-to-cell signalling within growing bacterial cellulose pellicles, Microbial Biotechnology, Vol: 12, Pages: 611-619, ISSN: 1751-7915
Bacterial cellulose is a strong and flexible biomaterial produced at high yields by Acetobacter species and has applications in health care, biotechnology and electronics. Naturally, bacterial cellulose grows as a large unstructured polymer network around the bacteria that produce it, and tools to enable these bacteria to respond to different locations are required to grow more complex structured materials. Here, we introduce engineered cell‐to‐cell communication into a bacterial cellulose‐producing strain of Komagataeibacter rhaeticus to enable different cells to detect their proximity within growing material and trigger differential gene expression in response. Using synthetic biology tools, we engineer Sender and Receiver strains of K. rhaeticus to produce and respond to the diffusible signalling molecule, acyl‐homoserine lactone. We demonstrate that communication can occur both within and between growing pellicles and use this in a boundary detection experiment, where spliced and joined pellicles sense and reveal their original boundary. This work sets the basis for synthetic cell‐to‐cell communication within bacterial cellulose and is an important step forward for pattern formation within engineered living materials.
Berry J, Gurung I, Anonsen JH, et al., 2019, Global biochemical and structural analysis of the type IV pilus from the Gram-positive bacterium Streptococcus sanguinis, Journal of Biological Chemistry, Vol: 294, Pages: 6796-6808, ISSN: 0021-9258
Type IV pili (Tfp) are functionally versatile filaments, widespread in prokaryotes, that belong to a large class of filamentous nanomachines known as type IV filaments (Tff). Although Tfp have been extensively studied in several Gram-negative pathogens where they function as key virulence factors, many aspects of their biology remain poorly understood. Here, we performed a global biochemical and structural analysis of Tfp in a recently emerged Gram-positive model, Streptococcus sanguinis. In particular, we focused on the five pilins and pilin-like proteins involved in Tfp biology in S. sanguinis. We found that the two major pilins, PilE1 and PilE2, (i) follow widely conserved principles for processing by the prepilin peptidase PilD and for assembly into filaments; (ii) display only one of the post-translational modifications frequently found in pilins, i.e. a methylated N-terminus; (iii) are found in the same hetero-polymeric filaments; and (iv) are not functionally equivalent. The 3D structure of PilE1, solved by NMR, revealed a classical pilin fold with a highly unusual flexible C-terminus. Intriguingly, PilE1 more closely resembles pseudopilins forming shorter Tff than bona fide Tfp-forming major pilins, underlining the evolutionary relatedness among different Tff. Finally, we show that S. sanguinis Tfp contain a low abundance of three additional proteins processed by PilD, the minor pilins PilA, PilB, and PilC. These findings provide the first global biochemical and structural picture of a Gram-positive Tfp and have fundamental implications for our understanding of a widespread class of filamentous nanomachines.
Goosens VJ, Busch A, Georgiadou M, et al., 2017, Reconstitution of a minimal machinery capable of assembling type IV pili, Proceedings of the National Academy of Sciences of the United States of America, Vol: 114, Pages: E4978-E4986, ISSN: 0027-8424
Type IV pili (Tfp), which are key virulence factors in many bacterial pathogens, define a large group of multipurpose filamentous nanomachines widespread in Bacteria and Archaea. Tfp biogenesis is a complex multistep process, which relies on macromolecular assemblies composed of 15 conserved proteins in model gram-negative species. To improve our limited understanding of the molecular mechanisms of filament assembly, we have used a synthetic biology approach to reconstitute, in a nonnative heterologous host, a minimal machinery capable of building Tfp. Here we show that eight synthetic genes are sufficient to promote filament assembly and that the corresponding proteins form a macromolecular complex at the cytoplasmic membrane, which we have purified and characterized biochemically. Our results contribute to a better mechanistic understanding of the assembly of remarkable dynamic filaments nearly ubiquitous in prokaryotes.
Neef J, Bongiorni C, Goosens VJ, et al., 2017, Intramembrane protease RasP boosts protein production in Bacillus, MICROBIAL CELL FACTORIES, Vol: 16, ISSN: 1475-2859
Goosens VJ, van Dijl JM, 2017, Twin-arginine protein translocation, Protein and Sugar Export and Assembly in Gram-positive Bacteria, Editors: Bagnoli, Rappuoli, Publisher: SPRINGER-VERLAG BERLIN, Pages: 69-94, ISBN: 978-3-319-56012-0
Twin-arginine protein translocation systems (Tat) translocate fully folded and co-factor-containing proteins across biological membranes. In this review, we focus on the Tat pathway of Gram-positive bacteria. The minimal Tat pathway is composed of two components, namely a TatA and TatC pair, which are often complemented with additional TatA-like proteins. We provide overviews of our current understanding of Tat pathway composition and mechanistic aspects related to Tat-dependent cargo protein translocation. This includes Tat pathway flexibility, requirements for the correct folding and incorporation of co-factors in cargo proteins and the functions of known cargo proteins. Tat pathways of several Gram-positive bacteria are discussed in detail, with emphasis on the Tat pathway of Bacillus subtilis. We discuss both shared and unique features of the different Gram-positive bacterial Tat pathways. Lastly, we highlight topics for future research on Tat, including the development of this protein transport pathway for the biotechnological secretion of high-value proteins and its potential applicability as an antimicrobial drug target in pathogens.
Goosens VJ, De-San-Eustaquio-Campillo A, Carballido-Lopez R, et al., 2015, A Tat menage a trois - The role of Bacillus subtilis TatAc in twin-arginine protein translocation, BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, Vol: 1853, Pages: 2745-2753, ISSN: 0167-4889
Goosens VJ, Monteferrante CG, van Dijl JM, 2014, The Tat system of Gram-positive bacteria, BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, Vol: 1843, Pages: 1698-1706, ISSN: 0167-4889
Goosens VJ, Monteferrante CG, van Dijl JM, 2014, Co-factor Insertion and Disulfide Bond Requirements for Twin-arginine Translocase-dependent Export of the Bacillus subtilis Rieske Protein QcrA, JOURNAL OF BIOLOGICAL CHEMISTRY, Vol: 289, Pages: 13124-13131
Moolla N, Goosens VJ, Kana BD, et al., 2014, The contribution of Nth and Nei DNA glycosylases to mutagenesis in Mycobacterium smegmatis, DNA REPAIR, Vol: 13, Pages: 32-41, ISSN: 1568-7864
Krishnappa L, Dreisbach A, Otto A, et al., 2013, Extracytoplasmic Proteases Determining the Cleavage and Release of Secreted Proteins, Lipoproteins, and Membrane Proteins in Bacillus subtilis, JOURNAL OF PROTEOME RESEARCH, Vol: 12, Pages: 4101-4110, ISSN: 1535-3893
Goosens VJ, Mars RAT, Akeroyd M, et al., 2013, Is Proteomics a Reliable Tool to Probe the Oxidative Folding of Bacterial Membrane Proteins?, ANTIOXIDANTS & REDOX SIGNALING, Vol: 18, Pages: 1159-1164, ISSN: 1523-0864
Goosens VJ, Otto A, Glasner C, et al., 2013, Novel Twin-Arginine Translocation Pathway-Dependent Phenotypes of Bacillus subtilis Unveiled by Quantitative Proteomics, JOURNAL OF PROTEOME RESEARCH, Vol: 12, Pages: 796-807, ISSN: 1535-3893
van der Ploeg R, Barnett JP, Vasisht N, et al., 2011, Salt Sensitivity of Minimal Twin Arginine Translocases, JOURNAL OF BIOLOGICAL CHEMISTRY, Vol: 286, Pages: 43759-43770, ISSN: 0021-9258
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