RESUMEN
CARD8 senses pathogen-associated protease activity and assembles a pyroptosis-inducing inflammasome, but detailed regulatory mechanisms have remained elusive. In this issue of Immunity, Sharif et al. use cryo-EM and biochemical assays to unveil how DPP9 sequesters the inflammasome-forming C-terminal fragment of CARD8 to suppress its activation.
Asunto(s)
Proteínas Adaptadoras de Señalización CARD , Inflamasomas , Proteínas Reguladoras de la Apoptosis/metabolismo , Proteínas Adaptadoras de Señalización CARD/genética , Proteínas Adaptadoras de Señalización CARD/metabolismo , Inflamasomas/metabolismo , Proteínas de Neoplasias/metabolismo , PiroptosisRESUMEN
RIG-I is a cytosolic receptor of viral RNA essential for the immune response to numerous RNA viruses. Accordingly, RIG-I must sensitively detect viral RNA yet tolerate abundant self-RNA species. The basic binding cleft and an aromatic amino acid of the RIG-I C-terminal domain(CTD) mediate high-affinity recognition of 5'triphosphorylated and 5'base-paired RNA(dsRNA). Here, we found that, while 5'unmodified hydroxyl(OH)-dsRNA demonstrated residual activation potential, 5'-monophosphate(5'p)-termini, present on most cellular RNAs, prevented RIG-I activation. Determination of CTD/dsRNA co-crystal structures and mutant activation studies revealed that the evolutionarily conserved I875 within the CTD sterically inhibits 5'p-dsRNA binding. RIG-I(I875A) was activated by both synthetic 5'p-dsRNA and endogenous long dsRNA within the polyA-rich fraction of total cellular RNA. RIG-I(I875A) specifically interacted with long, polyA-bearing, mitochondrial(mt) RNA, and depletion of mtRNA from total RNA abolished its activation. Altogether, our study demonstrates that avoidance of 5'p-RNA recognition is crucial to prevent mtRNA-triggered RIG-I-mediated autoinflammation.
Asunto(s)
Proteína 58 DEAD Box , Isoleucina , Receptores Inmunológicos , Proteína 58 DEAD Box/química , Proteína 58 DEAD Box/genética , Proteína 58 DEAD Box/metabolismo , Tolerancia Inmunológica , Isoleucina/genética , ARN Bicatenario/genética , ARN Mitocondrial/genética , ARN Mitocondrial/metabolismo , ARN Viral/genética , ARN Viral/metabolismo , Humanos , Receptores Inmunológicos/química , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismoRESUMEN
BACKGROUND: Our objective was to develop a three-dimensional high-resolution ultrasonic imaging technique to be utilized for in-vivo characterization of the ciliary body and the posterior iris. The benefit of this imaging in enhancing the quantification of the configurational changes in the ciliary body during accommodation is demonstrated. METHODS: Sequential ultrasound biomicroscopic images of the ciliary body region were obtained with a computer-controlled scanning device designed for use with a standard ultrasound biomicroscope for 3D imaging. Custom-made software allows online data collection, data analysis and 3D reconstruction in conjunction with commercially available VoxelView software. RESULTS: The three-dimensional presentation allows a close approximation of the ciliary muscle inside the ciliary body in vivo. We are able to distinguish and to analyze the changes in the muscle contour in different accommodation states. During accommodation a shift in the ciliary muscle center of gravity in a range of 0.04-0.26 mm (mean 0.13+/-0.06 mm) in the direction of the lens equator, with an interindividual variation and a small decrease with age, was observed. CONCLUSIONS: High-resolution ultrasound is a well established technique for in-vivo investigation of the anterior segment. Three-dimensional ultrasound biomicroscopy allows an assessment of the individual ciliary muscle activity in consideration of the ciliary processes. In combination with a contour analysis tool we improved the muscle contour determination during different accommodation states. The investigation showed an activity of the ciliary muscle in young volunteers as well as those of presbyopic age.