The Botrytis cinerea xylanase Xyn11A contributes to virulence with its necrotizing activity, not with its catalytic activity

<p>Abstract</p> <p>Background</p> <p>The <it>Botrytis cinerea </it>xylanase Xyn11A has been previously shown to be required for full virulence of this organism despite its poor contribution to the secreted xylanase activity and the low xylan content of <it>B. cinerea </it>hosts. Intriguingly, xylanases from other fungi have been shown to have the property, independent of the xylan degrading activity, to induce necrosis when applied to plant tissues, so we decided to test the hypothesis that secreted Xyn11A contributes to virulence by promoting the necrosis of the plant tissue surrounding the infection, therefore facilitating the growth of this necrotroph.</p> <p>Results</p> <p>We show here that Xyn11A has necrotizing activity on plants and that this capacity is conserved in site-directed mutants of the protein lacking the catalytic activity. Besides, Xyn11A contributes to the infection process with the necrotizing and not with the xylan hydrolyzing activity, as the catalytically-impaired Xyn11A variants were able to complement the lower virulence of the <it>xyn11A </it>mutant. The necrotizing activity was mapped to a 30-amino acids peptide in the protein surface, and this region was also shown to mediate binding to tobacco spheroplasts by itself.</p> <p>Conclusions</p> <p>The main contribution of the xylanase Xyn11A to the infection process of <it>B. cinerea </it>is to induce necrosis of the infected plant tissue. A conserved 30-amino acids region on the enzyme surface, away from the xylanase active site, is responsible for this effect and mediates binding to plant cells.</p

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Slow slip source characterized by lithological and geometric heterogeneity

Slow slip events (SSEs) accommodate a significant proportion of tectonic plate motion at subduction zones, yet little is known about the faults that actually host them. The shallow depth (<2 km) of well-documented SSEs at the Hikurangi subduction zone offshore New Zealand offers a unique opportunity to link geophysical imaging of the subduction zone with direct access to incoming material that represents the megathrust fault rocks hosting slow slip. Two recent International Ocean Discovery Program Expeditions sampled this incoming material before it is entrained immediately down-dip along the shallow plate interface. Drilling results, tied to regional seismic reflection images, reveal heterogeneous lithologies with highly variable physical properties entering the SSE source region. These observations suggest that SSEs and associated slow earthquake phenomena are promoted by lithological, mechanical, and frictional heterogeneity within the fault zone, enhanced by geometric complexity associated with subduction of rough crust

Expedition 372B/375 summary

Slow slip events (SSEs) at the northern Hikurangi subduction margin, New Zealand, are among the best-documented shallow SSEs on Earth. International Ocean Discovery Program Expeditions 372 and 375 were undertaken to investigate the processes and in situ conditions that underlie subduction zone SSEs at the northern Hikurangi Trough. We accomplished this goal by (1) coring and geophysical logging at four sites, including penetration of an active thrust fault (the Pāpaku fault) near the deformation front, the upper plate above the SSE source region, and the incoming sedimentary succession in the Hikurangi Trough and atop the Tūranganui Knoll seamount; and (2) installing borehole observatories in the Pāpaku fault and in the upper plate overlying the slow slip source region. Logging-while-drilling (LWD) data for this project were acquired as part of Expedition 372, and coring, wireline logging, and observatory installations were conducted during Expedition 375. Northern Hikurangi subduction margin SSEs recur every 1–2 y and thus provide an ideal opportunity to monitor deformation and associated changes in chemical and physical properties throughout the slow slip cycle. In situ measurements and sampling of material from the sedimentary section and oceanic basement of the subducting plate reveal the rock properties, composition, lithology, and structural character of material that is transported downdip into the SSE source region. A recent seafloor geodetic experiment raises the possibility that SSEs at northern Hikurangi may propagate to the trench, indicating that the shallow thrust fault (the Pāpaku fault) targeted during Expeditions 372 and 375 may also lie in the SSE rupture area and host a portion of the slip in these events. Hence, sampling and logging at this location provides insights into the composition, physical properties, and architecture of a shallow fault that may host slow slip. Expeditions 372 and 375 were designed to address three fundamental scientific objectives: Characterize the state and composition of the incoming plate and shallow fault near the trench, which comprise the protolith and initial conditions for fault zone rock at greater depth and which may itself host shallow slow slip; Characterize material properties, thermal regime, and stress conditions in the upper plate directly above the SSE source region; and Install observatories in the Pāpaku fault near the deformation front and in the upper plate above the SSE source to measure temporal variations in deformation, temperature, and fluid flow. The observatories will monitor volumetric strain (via pore pressure as a proxy) and the evolution of physical, hydrological, and chemical properties throughout the SSE cycle. Together, the coring, logging, and observatory data will test a suite of hypotheses about the fundamental mechanics and behavior of SSEs and their relationship to great earthquakes along the subduction interface

Revision of Coelastrella (Scenedesmaceae, Chlorophyta) and first register of this green coccoid microalga for continental Norway.

publishedVersio

SGIP1 modulates kinetics and interactions of the cannabinoid receptor 1 and G protein‐coupled receptor kinase 3 signalosome

International audienceCannabinoid receptor 1 (CB1R), a G protein-coupled receptor, plays a fundamental role in synaptic plasticity. Abnormal activity and deregulation of CB1R signaling result in a broad spectrum of pathological conditions. CB1R signaling is regulated by receptor desensitization including phosphorylation of residues within the intracellular C terminus by G protein-coupled receptor kinases (GRKs) that may lead to endocytosis. Furthermore, CB1R signaling is regulated by the protein Src homology 3-domain growth factor receptor-bound 2-like (SGIP1) that hinders receptor internalization, while enhancing CB1R association with β-arrestin. It has been postulated that phosphorylation of two clusters of serine/threonine residues, 425 SMGDS429 and 460 TMSVSTDTS468 , within the CB1R C-tail controls dynamics of the association between receptor and its interaction partners involved in desensitization. Several molecular determinants of these events are still not well understood. We hypothesized that the dynamics of these interactions are modulated by SGIP1. Using a panel of CB1Rs mutated in the aforementioned serine and threonine residues, together with an array of Bioluminescence energy transfer-based (BRET) sensors, we discovered that GRK3 forms complexes with Gβγ subunits of G proteins that largely independent of GRK3's interaction with CB1R. Furthermore, CB1R interacts only with activated GRK3. Interestingly, phosphorylation of two specific residues on CB1R triggers GRK3 dissociation from the desensitized receptor. SGIP1 increases the association of GRK3 with Gβγ subunits of G proteins, and with CB1R. Altogether, our data suggest that the CB1R signalosome complex is dynamically controlled by sequential phosphorylation of the receptor C-tail and is also modified by SGIP1