Crystal Structure of Thermotoga maritima α-Glucosidase AglA Defines a New Clan of NAD+-dependent Glycosidases

Glycoside hydrolase family 4 represents an unusual group of glucosidases with a requirement for NAD(+), divalent metal cations, and reducing conditions. The family is also unique in its inclusion of both alpha- and beta-specific enzymes. The alpha-glucosidase A, AglA, from Thermotoga maritima is a typical glycoside hydrolase family 4 enzyme, requiring NAD(+) and Mn2+ as well as strongly reducing conditions for activity. Here we present the crystal structure of the protein complexed with NAD(+) and maltose, refined at a resolution of 1.9 Angstrom. The NAD(+) is bound to a typical Rossman fold NAD(+)-binding site, and the nicotinamide moiety is localized close to the maltose substrate. Within the active site the conserved Cys-174 and surrounding histidines are positioned to play a role in the hydrolysis reaction. The electron density maps indicate that Cys-174 is oxidized to a sulfinic acid. Most likely, the strongly reducing conditions are necessary to reduce the oxidized cysteine side chain. Notably, the canonical set of catalytic acidic residues common to other glucosidases is not present in the active site. This, combined with a high structural homology to NAD-dependent dehydrogenases, suggests an unusual and possibly unique mechanism of action for a glycoside-hydrolyzing enzyme

Stress related epigenetic changes may explain opportunistic success in biological invasions in Antipode mussels

Different environmental factors could induce epigenetic changes, which are likely involved in the biological invasion process. Some of these factors are driven by humans as, for example, the pollution and deliberate or accidental introductions and others are due to natural conditions such as salinity. In this study, we have analysed the relationship between different stress factors: time in the new location, pollution and salinity with the methylation changes that could be involved in the invasive species tolerance to new environments. For this purpose, we have analysed two different mussels’ species, reciprocally introduced in antipode areas: the Mediterranean blue mussel Mytilus galloprovincialis and the New Zealand pygmy mussel Xenostrobus securis, widely recognized invaders outside their native distribution ranges. The demetylathion was higher in more stressed population, supporting the idea of epigenetic is involved in plasticity process. These results can open a new management protocols, using the epigenetic signals as potential pollution monitoring tool. We could use these epigenetic marks to recognise the invasive status in a population and determine potential biopollutants

Long time black hole evaporation with bounded Hawking flux

The long time behavior of an evaporating Schwarzschild black hole is studied exploiting that it can be described by an effective theory in 2D, a particular dilaton gravity model. A crucial technical ingredient is Izawa's result on consistent deformations of 2D BF theory, while the most relevant physical assumption is boundedness of the asymptotic matter flux during the whole evaporation process. An attractor solution, the endpoint of the evaporation process, is found. Its metric is flat. However, the behavior of the dilaton field is nontrivial: it is argued that during the final flicker a first order phase transition occurs from a linear to a constant dilaton vacuum, thereby emitting a shock wave with a total energy of a fraction of the Planck mass. Another fraction of the Planck mass may reside in a cold remnant. [Note: More detailed abstract in the paper]Comment: 34 pages, 6 figures, v2: included new references and 2 new footnotes; v3: mayor revisions (extended intro, included pedagogical example, rearranged presentation, extended discussion on information paradox, updated references); v4: updated refs. (+ new ones), added comments, mostly on dilaton evaporation, rewrote abstract (short for arXiv, long for journal), moved pedagogic sec. to ap

An action for the exact string black hole

A local action is constructed describing the exact string black hole discovered by Dijkgraaf, Verlinde and Verlinde in 1992. It turns out to be a special 2D Maxwell-dilaton gravity theory, linear in curvature and field strength. Two constants of motion exist: mass M>1, determined by the level k, and U(1)-charge Q>0, determined by the value of the dilaton at the origin. ADM mass, Hawking temperature T_H \propto \sqrt{1-1/M} and Bekenstein-Hawking entropy are derived and studied in detail. Winding/momentum mode duality implies the existence of a similar action, arising from a branch ambiguity, which describes the exact string naked singularity. In the strong coupling limit the solution dual to AdS_2 is found to be the 5D Schwarzschild black hole. Some applications to black hole thermodynamics and 2D string theory are discussed and generalizations - supersymmetric extension, coupling to matter and critical collapse, quantization - are pointed out.Comment: 41 pages, 2 eps figures, dedicated to Wolfgang Kummer on occasion of his Emeritierung; v2: added ref; v3: extended discussion in sections 3.2, 3.3 and at the end of 5.3 by adding 2 pages of clarifying text; updated refs; corrected typo

The Human Fungal Pathogen Cryptococcus neoformans Escapes Macrophages by a Phagosome Emptying Mechanism That Is Inhibited by Arp2/3 Complex-Mediated Actin Polymerisation

The lysis of infected cells by disease-causing microorganisms is an efficient but risky strategy for disseminated infection, as it exposes the pathogen to the full repertoire of the host's immune system. Cryptococcus neoformans is a widespread fungal pathogen that causes a fatal meningitis in HIV and other immunocompromised patients. Following intracellular growth, cryptococci are able to escape their host cells by a non-lytic expulsive mechanism that may contribute to the invasion of the central nervous system. Non-lytic escape is also exhibited by some bacterial pathogens and is likely to facilitate long-term avoidance of the host immune system during latency. Here we show that phagosomes containing intracellular cryptococci undergo repeated cycles of actin polymerisation. These actin ‘flashes’ occur in both murine and human macrophages and are dependent on classical WASP-Arp2/3 complex mediated actin filament nucleation. Three dimensional confocal imaging time lapse revealed that such flashes are highly dynamic actin cages that form around the phagosome. Using fluorescent dextran as a phagosome membrane integrity probe, we find that the non-lytic expulsion of Cryptococcus occurs through fusion of the phagosome and plasma membranes and that, prior to expulsion, 95% of phagosomes become permeabilised, an event that is immediately followed by an actin flash. By using pharmacological agents to modulate both actin dynamics and upstream signalling events, we show that flash occurrence is inversely related to cryptococcal expulsion, suggesting that flashes may act to temporarily inhibit expulsion from infected phagocytes. In conclusion, our data reveal the existence of a novel actin-dependent process on phagosomes containing cryptococci that acts as a potential block to expulsion of Cryptococcus and may have significant implications for the dissemination of, and CNS invasion by, this organism.\ud \u

Novel insights into host-fungal pathogen interactions derived from live-cell imaging

Acknowledgments The authors acknowledge funding from the Wellcome Trust (080088, 086827, 075470 and 099215) including a Wellcome Trust Strategic Award for Medical Mycology and Fungal Immunology 097377 and FP7-2007–2013 grant agreement HEALTH-F2-2010-260338–ALLFUN to NARG.Peer reviewedPublisher PD

Thermodynamics of Black Holes in Two (and Higher) Dimensions

A comprehensive treatment of black hole thermodynamics in two-dimensional dilaton gravity is presented. We derive an improved action for these theories and construct the Euclidean path integral. An essentially unique boundary counterterm renders the improved action finite on-shell, and its variational properties guarantee that the path integral has a well-defined semi-classical limit. We give a detailed discussion of the canonical ensemble described by the Euclidean partition function, and examine various issues related to stability. Numerous examples are provided, including black hole backgrounds that appear in two dimensional solutions of string theory. We show that the Exact String Black Hole is one of the rare cases that admits a consistent thermodynamics without the need for an external thermal reservoir. Our approach can also be applied to certain higher-dimensional black holes, such as Schwarzschild-AdS, Reissner-Nordstrom, and BTZ.Comment: 63 pages, 3 pdf figures, v2: added reference