CYRI/FAM49B negatively regulates RAC1-driven cytoskeletal remodelling and protects against bacterial infection

Yuki, Kyoko E.; Marei, Hadir; Fiškin, Evgenij; Eva, Megan M.; Gopal, Angelica A.; Schwartzentruber, Jeremy; Majewski, Jacek; Cellier, Mathieu; Mandl, Judith N.; Vidal, Silvia M.; Malo, Danielle; Đikić, Ivan

doi:10.1038/s41564-019-0484-8

Cited by 42 publications

(42 citation statements)

References 61 publications

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“…Two CYFIP isoforms exist, CYFIP1 and CYFIP2, which both contain a DUF1394 domain (residues 59-301; human numbering) that is also shared by CYRI proteins. Despite a relatively low sequence identity of 21% between human CYRI-B and the human CYFIP1 DUF1394 domain, the two regions are predicted to be structurally homologous (Yuki et al, 2019). Comparison of the CYFIP1 DUF1394 domain (PDB entry 3p8c) with our structure of CYRI-B confirms this prediction (Fig.…”

Section: Similarity To Cyfip1 From the Wave Regulatory Complex (Wrc)supporting

confidence: 60%

“…After optimization and screening of multiple crystals, we were able to obtain diffraction data at 2.37 Å resolution. We attempted to solve the structure by molecular replacement using the DUF1394 domain of CYFIP1 (PDB entry 3p8c; Chen et al, 2010), as it shares 21% sequence identity with whale shark CYRI-B and is predicted to be structurally similar (Yuki et al, 2019). However, these efforts were unsuccessful.…”

Section: Resultsmentioning

confidence: 99%

“…CYRI-B binding has been reported to block various Rac1-dependent signalling cascades in the cell, thereby controlling multiple critical cellular functions including T-cell activation (Shang et al, 2018), membrane protrusion, chemotaxis and cell migration (Fort et al, 2018;Whitelaw et al, 2019). By negatively regulating Rac1 signalling, CYRI-B also reduces the entry of several intracellular bacterial pathogens into both phagocytic and non-phagocytic host cells, and indeed is targeted for ubiquitin-mediated destruction by the action of an injected Salmonella virulence protein (Yuki et al, 2019). CYRI-B has also been suggested to play a role in certain cancers.…”

Section: Introductionmentioning

confidence: 99%

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Structure of CYRI-B (FAM49B), a key regulator of cellular actin assembly

Kaplan

Stone

Hume

et al. 2020

Acta Crystallogr D Struct Biol

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In eukaryotes, numerous fundamental processes are controlled by the WAVE regulatory complex (WRC) that regulates cellular actin polymerization, crucial for cell motility, cell–cell adhesion and epithelial differentiation. Actin assembly is triggered by interaction of the small GTPase Rac1 with CYFIP1, a key component of the WRC. Previously known as FAM49B, CYRI-B is a protein that is highly conserved across the Eukaryota and has recently been revealed to be a key regulator of Rac1 activity. Mutation of CYRI-B or alteration of its expression therefore leads to altered actin nucleation dynamics, with impacts on lamellipodia formation, cell migration and infection by intracellular pathogens. In addition, knockdown of CYRI-B expression in cancer cell lines results in accelerated cell proliferation and invasiveness. Here, the structure of Rhincodon typus (whale shark) CYRI-B is presented, which is the first to be reported of any CYRI family member. Solved by X-ray crystallography, the structure reveals that CYRI-B comprises three distinct α-helical subdomains and is highly structurally related to a conserved domain present in CYFIP proteins. The work presented here establishes a template towards a better understanding of CYRI-B biological function.

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Section: Similarity To Cyfip1 From the Wave Regulatory Complex (Wrc)supporting

confidence: 60%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structure of CYRI-B (FAM49B), a key regulator of cellular actin assembly

Kaplan

Stone

Hume

et al. 2020

Acta Crystallogr D Struct Biol

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“…It is possible therefore that both WAVE truncation and activating CYFIP2 mutations can similarly sequester away active Rac, thereby suppressing Rac functions and its associated effector pathways. This could work in a fashion comparable to what has recently been suggested for the Rac binding protein FAM49B [29][30][31]. Such a scenario would also fit the observation that certain missense mutations in Rac1 leading to ID cause its loss of function [22], as well as the model that signaling of the "Rac1-antagonist" RhoA is typically upregulated and Rac signaling suppressed in ID [32].…”

Section: Discussionsupporting

confidence: 62%

Molecular Dissection of Neurodevelopmental Disorder-Causing Mutations in CYFIP2

Schaks

Reinke

Witke

et al. 2020

Cells

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Actin remodeling is frequently regulated by antagonistic activities driving protrusion and contraction downstream of Rac and Rho small GTPases, respectively. WAVE regulatory complex (WRC), which primarily operates downstream of Rac, plays pivotal roles in neuronal morphogenesis. Recently, two independent studies described de novo mutations in the CYFIP2 subunit of WRC, which caused intellectual disability (ID) in humans. Although mutations had been proposed to effect WRC activation, no experimental evidence for this was provided. Here, we made use of CRISPR/Cas9-engineered B16-F1 cell lines that were reconstituted with ID-causing CYFIP variants in different experimental contexts. Almost all CYFIP2-derived mutations (7 out of 8) promoted WRC activation, but to variable extent and with at least two independent mechanisms. The majority of mutations occurs in a conserved WAVE-binding region, required for WRC transinhibition. One mutation is positioned closely adjacent to the Rac-binding A site and appears to ease Rac-mediated WRC activation. As opposed to these gain-of-function mutations, a truncating mutant represented a loss-of-function variant and failed to interact with WRC components. Collectively, our data show that explored CYFIP2 mutations frequently, but not always, coincide with WRC activation and suggest that normal brain development requires a delicate and precisely tuned balance of neuronal WRC activity.

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“…Mice were infected by oral gavage with 200 l of a 10-fold dilution of the overnight culture containing 2 ϫ 10 7 to 3 ϫ 10 7 CFU of Salmonella spp. The infectious doses were verified by the plating of serial dilutions on Trypticase soy agar (48,49). Mice infected orally were first deprived of food for 4 h before infection with Salmonella enterica.…”

Section: Methodsmentioning

confidence: 99%

Combining Whole-Genome Sequencing and Multimodel Phenotyping To Identify Genetic Predictors of Salmonella Virulence

Crouse

Schramm

Emond-Rhéault

et al. 2020

mSphere

Self Cite

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Salmonella comprises more than 2,600 serovars. Very few environmental and uncommon serovars have been characterized for their potential role in virulence and human infections. A complementary in vitro and in vivo systematic high-throughput analysis of virulence was used to elucidate the association between genetic and phenotypic variations across Salmonella isolates. The goal was to develop a strategy for the classification of isolates as a benchmark and predict virulence levels of isolates. Thirty-five phylogenetically distant strains of unknown virulence were selected from the Salmonella Foodborne Syst-OMICS (SalFoS) collection, representing 34 different serovars isolated from various sources. Isolates were evaluated for virulence in 4 complementary models of infection to compare virulence traits with the genomics data, including interactions with human intestinal epithelial cells, human macrophages, and amoeba. In vivo testing was conducted using the mouse model of Salmonella systemic infection. Significant correlations were identified between the different models. We identified a collection of novel hypothetical and conserved proteins associated with isolates that generate a high burden. We also showed that blind prediction of virulence of 33 additional strains based on the pan-genome was high in the mouse model of systemic infection (82% agreement) and in the human epithelial cell model (74% agreement). These complementary approaches enabled us to define virulence potential in different isolates and present a novel strategy for risk assessment of specific strains and for better monitoring and source tracking during outbreaks. IMPORTANCE Salmonella species are bacteria that are a major source of foodborne disease through contamination of a diversity of foods, including meat, eggs, fruits, nuts, and vegetables. More than 2,600 different Salmonella enterica serovars have been identified, and only a few of them are associated with illness in humans. Despite the fact that they are genetically closely related, there is enormous variation in the virulence of different isolates of Salmonella enterica. Identification of foodborne pathogens is a lengthy process based on microbiological, biochemical, and immunological methods. Here, we worked toward new ways of integrating whole-genome sequencing (WGS) approaches into food safety practices. We used WGS to build associations between virulence and genetic diversity within 83 Salmonella isolates representing 77 different Salmonella serovars. Our work demonstrates the potential of combining a genomics approach and virulence tests to improve the diagnostics and assess risk of human illness associated with specific Salmonella isolates.

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CYRI/FAM49B negatively regulates RAC1-driven cytoskeletal remodelling and protects against bacterial infection

Cited by 42 publications

References 61 publications

Structure of CYRI-B (FAM49B), a key regulator of cellular actin assembly

Structure of CYRI-B (FAM49B), a key regulator of cellular actin assembly

Molecular Dissection of Neurodevelopmental Disorder-Causing Mutations in CYFIP2

Combining Whole-Genome Sequencing and Multimodel Phenotyping To Identify Genetic Predictors of Salmonella Virulence

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