Combined Crystal Structure of a Type I Cohesin

Cameron, Kate; Weinstein, Jonathan; Zhivin, Olga; Bule, Pedro; Fleishman, Sarel J.; Alves, Victor D.; Gilbert, Harry J.; Ferreira, L.M.A.; Fontes, Carlos M. G. A.; Bayer, Edward A.; Najmudin, Shabir

doi:10.1074/jbc.m115.653303

Cited by 10 publications

(5 citation statements)

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“…Previous research suggested that a dual-binding mode of the dockerins would result in increased flexibility characteristics of the catalytic subunits [ 67 , 68 ], it was reported previously that the type I ScaA dockerin has a dual-binding mode in the recognition of the B. cellulosolvens type I cohesins [ 69 ]. The additional type I dockerins reported here share similar binding preferences and sequence similarity.…”

Section: Discussionmentioning

confidence: 99%

Unique organization and unprecedented diversity of the Bacteroides (Pseudobacteroides) cellulosolvens cellulosome system

Zhivin

Dassa

Moraïs

et al. 2017

Biotechnol Biofuels

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Background (Pseudo) Bacteroides cellulosolvens is an anaerobic, mesophilic, cellulolytic, cellulosome-producing clostridial bacterium capable of utilizing cellulose and cellobiose as carbon sources. Recently, we sequenced the B. cellulosolvens genome, and subsequent comprehensive bioinformatic analysis, herein reported, revealed an unprecedented number of cellulosome-related components, including 78 cohesin modules scattered among 31 scaffoldins and more than 200 dockerin-bearing ORFs. In terms of numbers, the B. cellulosolvens cellulosome system represents the most intricate, compositionally diverse cellulosome system yet known in nature.ResultsThe organization of the B. cellulosolvens cellulosome is unique compared to previously described cellulosome systems. In contrast to all other known cellulosomes, the cohesin types are reversed for all scaffoldins i.e., the type II cohesins are located on the enzyme-integrating primary scaffoldin, whereas the type I cohesins are located on the anchoring scaffoldins. Many of the type II dockerin-bearing ORFs include X60 modules, which are known to stabilize type II cohesin–dockerin interactions. In the present work, we focused on revealing the architectural arrangement of cellulosome structure in this bacterium by examining numerous interactions between the various cohesin and dockerin modules. In total, we cloned and expressed 43 representative cohesins and 27 dockerins. The results revealed various possible architectures of cell-anchored and cell-free cellulosomes, which serve to assemble distinctive cellulosome types via three distinct cohesin–dockerin specificities: type I, type II, and a novel-type designated R (distinct from type III interactions, predominant in ruminococcal cellulosomes).ConclusionsThe results of this study provide novel insight into the architecture and function of the most intricate and extensive cellulosomal system known today, thereby extending significantly our overall knowledge base of cellulosome systems and their components. The robust cellulosome system of B. cellulosolvens, with its unique binding specificities and reversal of cohesin–dockerin types, has served to amend our view of the cellulosome paradigm. Revealing new cellulosomal interactions and arrangements is critical for designing high-efficiency artificial cellulosomes for conversion of plant-derived cellulosic biomass towards improved production of biofuels.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0898-6) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Unique organization and unprecedented diversity of the Bacteroides (Pseudobacteroides) cellulosolvens cellulosome system

Zhivin

Dassa

Moraïs

et al. 2017

Biotechnol Biofuels

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“…Three other PPIs showed no leakiness in the absence of one of the interaction partners, and at the same time showed a significant induction of SEAP when both parts were expressed in the same cell to reconstitute a functional TF ( Figure 1 c,d). These included (i) cohesin-dockerin proteins from Acetivibrio cellulolyticus (Coh(Ac)/Doc(Ac)), 21 (ii) antiparallel leucine zipper (Zip – /Zip + ) 22 domains from an engineered, bipartite TF used for gene control in Caenorhabditis elegans and (iii) the SunTag system (scFv(G4)/G4) 23 based on a single chain variable fragment of an antibody that specifically binds a yeast-derived 19 amino acid peptide. We observed no crosstalk between the different parts of the protein–protein interactions, which is required for their further assembly to multipartite TFs ( Figure 1 e).…”

Section: Resultsmentioning

confidence: 99%

Design of Multipartite Transcription Factors for Multiplexed Logic Genome Integration Control in Mammalian Cells

Ausländer

Auslander

et al. 2020

ACS Synth. Biol.

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Synthetic biology relies on rapid and efficient methods to stably integrate DNA payloads encoding for synthetic biological systems into the genome of living cells. The size of designed biological systems increases with their complexity, and novel methods are needed that enable efficient and simultaneous integration of multiple payloads into single cells. By assembling natural and synthetic protein–protein dimerization domains, we have engineered a set of multipartite transcription factors for driving heterologous target gene expression. With the distribution of single parts of multipartite transcription factors on piggyback transposon-based donor plasmids, we have created a logic genome integration control (LOGIC) system that allows for efficient one-step selection of stable mammalian cell lines with up to three plasmids. LOGIC significantly enhances the efficiency of multiplexed payload integration in mammalian cells compared to traditional cotransfection and may advance cell line engineering in synthetic biology and biotechnology.

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“…In addition, recent reports revealed that the attachment of cellulosomes to the P . cellulosolvens 21 and A . cellulolyticus cell surface is also mediated by Docs that display a dual-binding mode 17 , 22 .…”

Section: Discussionmentioning

confidence: 99%

Assembly of Ruminococcus flavefaciens cellulosome revealed by structures of two cohesin-dockerin complexes

Bule¹,

Alves²,

Israeli-Ruimy

et al. 2017

Sci Rep

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AbtractCellulosomes are sophisticated multi-enzymatic nanomachines produced by anaerobes to effectively deconstruct plant structural carbohydrates. Cellulosome assembly involves the binding of enzyme-borne dockerins (Doc) to repeated cohesin (Coh) modules located in a non-catalytic scaffoldin. Docs appended to cellulosomal enzymes generally present two similar Coh-binding interfaces supporting a dual-binding mode, which may confer increased positional adjustment of the different complex components. Ruminococcus flavefaciens’ cellulosome is assembled from a repertoire of 223 Doc-containing proteins classified into 6 groups. Recent studies revealed that Docs of groups 3 and 6 are recruited to the cellulosome via a single-binding mode mechanism with an adaptor scaffoldin. To investigate the extent to which the single-binding mode contributes to the assembly of R. flavefaciens cellulosome, the structures of two group 1 Docs bound to Cohs of primary (ScaA) and adaptor (ScaB) scaffoldins were solved. The data revealed that group 1 Docs display a conserved mechanism of Coh recognition involving a single-binding mode. Therefore, in contrast to all cellulosomes described to date, the assembly of R. flavefaciens cellulosome involves single but not dual-binding mode Docs. Thus, this work reveals a novel mechanism of cellulosome assembly and challenges the ubiquitous implication of the dual-binding mode in the acquisition of cellulosome flexibility.

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Combined Crystal Structure of a Type I Cohesin

Cited by 10 publications

References 38 publications

Unique organization and unprecedented diversity of the Bacteroides (Pseudobacteroides) cellulosolvens cellulosome system

Unique organization and unprecedented diversity of the Bacteroides (Pseudobacteroides) cellulosolvens cellulosome system

Design of Multipartite Transcription Factors for Multiplexed Logic Genome Integration Control in Mammalian Cells

Assembly of Ruminococcus flavefaciens cellulosome revealed by structures of two cohesin-dockerin complexes

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