RESUMEN
The type VI secretion system (T6SS) is deployed by numerous Gram-negative bacteria to deliver toxic effectors into neighbouring cells. The genome of Pantoea agglomerans pv. betae (Pab) phytopathogenic bacteria contains a gene cluster (T6SS1) predicted to encode a complete T6SS. Using secretion and competition assays, we found that T6SS1 in Pab is a functional antibacterial system that allows this pathogen to outcompete rival plant-associated bacteria found in its natural environment. Computational analysis of the T6SS1 gene cluster revealed that antibacterial effector and immunity proteins are encoded within three genomic islands that also harbour arrays of orphan immunity genes or toxin and immunity cassettes. Functional analyses indicated that VgrG, a specialized antibacterial effector, contains a C-terminal catalytically active glucosaminidase domain that is used to degrade prey peptidoglycan. Moreover, we confirmed that a bicistronic unit at the end of the T6SS1 cluster encodes a novel antibacterial T6SS effector and immunity pair. Together, these results demonstrate that Pab T6SS1 is an antibacterial system delivering a lysozyme-like effector to eliminate competitors, and indicate that this bacterium contains additional novel T6SS effectors.
Asunto(s)
Pantoea , Sistemas de Secreción Tipo VI , Antibacterianos/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Hexosaminidasas , Muramidasa/genética , Pantoea/genética , Peptidoglicano , Sistemas de Secreción Tipo VI/genética , Sistemas de Secreción Tipo VI/metabolismoRESUMEN
Gram-negative bacteria use type VI secretion systems (T6SSs) to deliver toxic effector proteins into neighboring cells. Cargo effectors are secreted by binding noncovalently to the T6SS apparatus. Occasionally, effector secretion is assisted by an adaptor protein, although the adaptor itself is not secreted. Here, we report a new T6SS secretion mechanism, in which an effector and a co-effector are secreted together. Specifically, we identify a novel periplasm-targeting effector that is secreted together with its co-effector, which contains a MIX (marker for type sIX effector) domain previously reported only in polymorphic toxins. The effector and co-effector directly interact, and they are dependent on each other for secretion. We term this new secretion mechanism "a binary effector module," and we show that it is widely distributed in marine bacteria.
Asunto(s)
Sistemas de Secreción Tipo VI , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Transporte Biológico , Sistemas de Secreción Tipo VI/genética , Sistemas de Secreción Tipo VI/metabolismoRESUMEN
Vibrionaceae is a widespread family of aquatic bacteria that includes emerging pathogens and symbionts. Many Vibrionaceae harbor a type VI secretion system (T6SS), which is a secretion apparatus used to deliver toxins, termed effectors, into neighboring cells. T6SSs mediate both antibacterial and anti-eukaryotic activities. Notably, antibacterial effectors are encoded together with a gene that encodes a cognate immunity protein so as to antagonize the toxicity of the effector. The MIX (Marker for type sIX effectors) domain has been previously defined as a marker of T6SS effectors carrying polymorphic C-terminal toxins. Here, we set out to identify the Vibrionaceae MIX-effector repertoire and to analyze the various toxin domains they carry. We used a computational approach to search for the MIX-effectors in the Vibrionaceae genomes, and grouped them into clusters based on the C-terminal toxin domains. We classified MIX-effectors as either antibacterial or anti-eukaryotic, based on the presence or absence of adjacent putative immunity genes, respectively. Antibacterial MIX-effectors carrying pore-forming, phospholipase, nuclease, peptidoglycan hydrolase, and protease activities were found. Furthermore, we uncovered novel virulence MIX-effectors. These are encoded by "professional MIXologist" strains that employ a cocktail of antibacterial and anti-eukaryotic MIX-effectors. Our findings suggest that certain Vibrionaceae adapted their antibacterial T6SS to mediate interactions with eukaryotic hosts or predators.
Asunto(s)
Antibacterianos/toxicidad , Organismos Acuáticos/metabolismo , Proteínas Bacterianas/toxicidad , Eucariontes/fisiología , Sistemas de Secreción Tipo VI/toxicidad , Vibrionaceae/metabolismo , Antibacterianos/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Biología Computacional , Interacciones Microbiota-Huesped/fisiología , Dominios Proteicos/genética , Sistemas de Secreción Tipo VI/genética , Sistemas de Secreción Tipo VI/metabolismo , Vibrionaceae/genética , Factores de Virulencia/genética , Factores de Virulencia/metabolismo , Factores de Virulencia/toxicidadRESUMEN
BACKGROUND: Although characteristics such as heart rate (HR) and blood pressure (BP) are commonly reported in studies of the relationship between fatigue and cardiac functioning, few reports examine how cardiac function parameters such as cardiac output (CO) and stroke volume (SV) relate to fatigue. This study examined the relationship between self-reported fatigue and hemodynamic functioning at rest and in response to a public speaking stressor in healthy individuals. METHODS: A total of 142 individuals participated in this study. Subjects were placed in low-, moderate-, or high-fatigue groups based on their Profile of Moods State fatigue scale. Heart rate, SV, and CO were determined using impedance cardiography at rest and during a speaking stressor. Stroke volume and CO values were converted to stroke index (SI) and cardiac index (CI) by adjusting for body surface area. Data were analyzed with hierarchical regression analysis and a 3 (group) x 3 (stress period) mixed model analysis of variance. RESULTS: At rest, fatigue was not associated with BP or HR but was significantly associated with decreased CI (P < .001; 95% confidence interval, -0.046 to -0.014) and stroke index (SI) (P = .002; 95% confidence interval -0.664 to -0.151), even after controlling for demographic variables and depressive symptoms. Heart rate and BP increased, as expected, from baseline to preparation to speaking stressor (F (1,124) = 118.6 and F (1,122) = 46.450, respectively) (P < .001 for both). More interestingly, there were effects on SI and CI of fatigue (P<.03 for both) and stress (P<.03 for both); high-fatigue individuals had lower SI and CI levels than moderate- and low-fatigue individuals both at rest and in response to the stressor. CONCLUSION: This study demonstrates that fatigue complaints may have hemodynamic correlates even in ostensibly healthy individuals.
Asunto(s)
Gasto Cardíaco/fisiología , Fatiga/fisiopatología , Frecuencia Cardíaca/fisiología , Volumen Sistólico/fisiología , Adulto , Presión Sanguínea/fisiología , Cardiografía de Impedancia , Femenino , Humanos , Masculino , Análisis de Regresión , Descanso/fisiología , Estrés Psicológico/fisiopatologíaRESUMEN
The basolateral (BLA) and medial nucleus (MeA) of the amygdala participate in the modulation of unconditioned fear induced by predator odor. However, the specific role of these amygdalar nuclei in predator odor-induced fear memory is not known. Therefore, fiber-sparing lesions or temporary inactivation of the BLA or MeA were made either prior to or after exposure to cat odor, and conditioned contextual fear behavior was examined the next day. BLA and MeA lesions produced significant reductions in cat odor-induced unconditioned and conditioned fear-related behavior. In addition, temporary pharmacological neural inactivation methods occurring after exposure to cat odor revealed subtle behavioral alterations indicative of a role of the BLA in fear memory consolidation but not memory retrieval. In contrast, the MeA appears to play a specific role in retrieval but not consolidation. Results show that the BLA participates in the conditioned and unconditioned cat odor stimulus association that underlies fear memory, underscore a novel role of the MeA in predator odor contextual conditioning, and demonstrate different roles of the BLA and MeA in modulating consolidation and retrieval of predator odor fear memory.