HDAC6 rescues neurodegeneration and provides an essential link between autophagy and the UPS

A prominent feature of late-onset neurodegenerative diseases is accumulation of misfolded protein in vulnerable neurons. When levels of misfolded protein overwhelm degradative pathways, the result is cellular toxicity and neurodegeneration. Cellular mechanisms for degrading misfolded protein include the ubiquitin-proteasome system (UPS), the main non-lysosomal degradative pathway for ubiquitinated proteins, and autophagy, a lysosome-mediated degradative pathway. The UPS and autophagy have long been viewed as complementary degradation systems with no point of intersection. This view has been challenged by two observations suggesting an apparent interaction: impairment of the UPS induces autophagy in vitro, and conditional knockout of autophagy in the mouse brain leads to neurodegeneration with ubiquitin-positive pathology. It is not known whether autophagy is strictly a parallel degradation system, or whether it is a compensatory degradation system when the UPS is impaired; furthermore, if there is a compensatory interaction between these systems, the molecular link is not known. Here we show that autophagy acts as a compensatory degradation system when the UPS is impaired in Drosophila melanogaster, and that histone deacetylase 6 (HDAC6), a microtubule-associated deacetylase that interacts with polyubiquitinated proteins, is an essential mechanistic link in this compensatory interaction. We found that compensatory autophagy was induced in response to mutations affecting the proteasome and in response to UPS impairment in a fly model of the neurodegenerative disease spinobulbar muscular atrophy. Autophagy compensated for impaired UPS function in an HDAC6-dependent manner. Furthermore, expression of HDAC6 was sufficient to rescue degeneration associated with UPS dysfunction in vivo in an autophagy-dependent manner. This study suggests that impairment of autophagy (for example, associated with ageing or genetic variation) might predispose to neurodegeneration. Morover, these findings suggest that it may be possible to intervene in neurodegeneration by augmenting HDAC6 to enhance autophagy.     

ATP-induced helicase slippage reveals highly coordinated subunits

Helicases are vital enzymes that carry out strand separation of duplex nucleic acids during replication, repair and recombination. Bacteriophage T7 gene product 4 is a model hexameric helicase that has been observed to use dTTP, but not ATP, to unwind double-stranded (ds)DNA as it translocates from 5' to 3' along single-stranded (ss)DNA. Whether and how different subunits of the helicase coordinate their chemo-mechanical activities and DNA binding during translocation is still under debate. Here we address this question using a single-molecule approach to monitor helicase unwinding. We found that T7 helicase does in fact unwind dsDNA in the presence of ATP and that the unwinding rate is even faster than that with dTTP. However, unwinding traces showed a remarkable sawtooth pattern where processive unwinding was repeatedly interrupted by sudden slippage events, ultimately preventing unwinding over a substantial distance. This behaviour was not observed with dTTP alone and was greatly reduced when ATP solution was supplemented with a small amount of dTTP. These findings presented an opportunity to use nucleotide mixtures to investigate helicase subunit coordination. We found that T7 helicase binds and hydrolyses ATP and dTTP by competitive kinetics such that the unwinding rate is dictated simply by their respective maximum rates V(max), Michaelis constants K(M) and concentrations. In contrast, processivity does not follow a simple competitive behaviour and shows a cooperative dependence on nucleotide concentrations. This does not agree with an uncoordinated mechanism where each subunit functions independently, but supports a model where nearly all subunits coordinate their chemo-mechanical activities and DNA binding. Our data indicate that only one subunit at a time can accept a nucleotide while other subunits are nucleotide-ligated and thus they interact with the DNA to ensure processivity. Such subunit coordination may be general to many ring-shaped helicases and reveals a potential mechanism for regulation of DNA unwinding during replication.     

Analysis of the human protein interactome and comparison with yeast, worm and fly interaction datasets

We present the first analysis of the human proteome with regard to interactions between proteins. We also compare the human interactome with the available interaction datasets from yeast (Saccharomyces cerevisiae), worm (Caenorhabditis elegans) and fly (Drosophila melanogaster). Of >70,000 binary interactions, only 42 were common to human, worm and fly, and only 16 were common to all four datasets. An additional 36 interactions were common to fly and worm but were not observed in humans, although a coimmunoprecipitation assay showed that 9 of the interactions do occur in humans. A re-examination of the connectivity of essential genes in yeast and humans indicated that the available data do not support the presumption that the number of interaction partners can accurately predict whether a gene is essential. Finally, we found that proteins encoded by genes mutated in inherited genetic disorders are likely to interact with proteins known to cause similar disorders, suggesting the existence of disease subnetworks. The human interaction map constructed from our analysis should facilitate an integrative systems biology approach to elucidating the cellular networks that contribute to health and disease states.     

Effect of electroslag remelting and homogenization on hydrogen flaking in AMS-4340 ultra-high-strength steels

Hydrogen flakes and elemental segregation are the main causes of steel rejection. To eliminate hydrogen flaking, the present study focuses on the manufacture of AMS-4340 ultra-high-strength steel through an alternate route. AMS-4340 was prepared using three different processing routes. The primary processing route consisted of melting in an electric arc furnace, refining in a ladle refining furnace, and vacuum degassing. After primary processing, the heat processes(D1, D2, and D3) were cast into cylindrical electrodes. For secondary processing, electroslag remelting(ESR) was carried out on the primary heats to obtain four secondary heats: E1, E2, E3, and E4. Homogenization of ingots E1, E2, E3, and E4 was carried out at 1220°C for 14, 12, 12, and 30 h, respectively, followed by an antiflaking treatment at 680°C and air cooling. In addition, the semi-finished ESR ingot E4 was again homogenized at 1220°C for 6–8 h and a second antiflaking treatment was performed at 680°C for 130 h followed by air cooling. The chemical segregation of each heat was monitored through a spectroscopy technique. The least segregation was observed for heat E4. Macrostructure examination revealed the presence of hydrogen flakes in heats E1, E2, and E3, whereas no hydrogen flakes were observed in heat E4. Ultrasonic testing revealed no internal defects in heat E4, whereas internal defects were observed in the other heats. A grain size investigation revealed a finer grain size for E4 compared with those for the other heats. Steel produced in heat E4 also exhibited superior mechanical properties. Therefore, the processing route used for heat E4 can be used to manufacture an AMS-4340 ultra-high-strength steel with superior properties compared with those of AMS-4340 prepared by the other investigated routes.     

等离子喷涂铁基非晶/晶体复合涂层的工艺–结构–性能关系

本研究试图通过优化等离子喷涂参数来开发一种Fe基非晶/晶体涂层,该涂层主要成分来自一种贫乏的铁基合金（Fe_92.6C_3.5P_1.4Si₂Mn_0.5）。这种合金是钢铁厂高炉产出的生铁剩余废料。为了经济有效地重新利用这种残留物,这种合金在合成时对成分进行了最少的修改。同时,本研究还探讨了涂层的结构、机械、腐蚀和磨损性能对喷涂参数（等离子功率、主气体流速、送粉速度和间隔距离）的依赖性。X射线衍射表明,在最优的喷涂参数下沉积的涂层存在无定形/晶体相。在较低等离子功率和最高气体流速下沉积的涂层表现出更好的密度、硬度和耐磨性。所有涂层都表现出良好的耐腐蚀性（腐蚀环境:3.5wt% NaCl 溶液）。机械、磨损和摩擦学研究表明,单一的工艺参数优化无法提供良好的涂层性能;相反,所有工艺参数在优化涂层性能都具有独一无二的作用,它们主要通过控制飞行中的颗粒温度和速度分布,以及熔滴撞击基材之前的冷却模式来控制涂层性能。     

Imaging the Indian subcontinent beneath the Himalaya

The rocks of the Indian subcontinent are last seen south of the Ganges before they plunge beneath the Himalaya and the Tibetan plateau. They are next glimpsed in seismic reflection profiles deep beneath southern Tibet, yet the surface seen there has been modified by processes within the Himalaya that have consumed parts of the upper Indian crust and converted them into Himalayan rocks. The geometry of the partly dismantled Indian plate as it passes through the Himalayan process zone has hitherto eluded imaging. Here we report seismic images both of the decollement at the base of the Himalaya and of the Moho (the boundary between crust and mantle) at the base of the Indian crust. A significant finding is that strong seismic anisotropy develops above the decollement in response to shear processes that are taken up as slip in great earthquakes at shallower depths. North of the Himalaya, the lower Indian crust is characterized by a high-velocity region consistent with the formation of eclogite, a high-density material whose presence affects the dynamics of the Tibetan plateau.     

Radioimmunoassay of progesterone in uterine flushings of buffalo (Bubalus bubalis)

The concentration of progesterone as determined by radioimmunoassay varied in accordance with the phase of ovarian activity.     

A new ester glucoside from the bark ofTecomella undulata

Zusammenfassung Isolierung und Strukturaufklärung eines neuen Esterglukosides aus der Rinde vonTecomella undulata.