Heterochromatin protein 1 modifies mammalian PEV in a dose- and chromosomal-context-dependent manner

Locus control regions (LCRs) are gene regulatory elements in mammals that can overcome the highly repressive effects normally associated with heterochromatic transgene locations (for example the centromere) in mice. Deletion of essential LCR sequences renders the cognate gene susceptible to this form of repression, so a proportion of the cells from transgenic mice that would normally express the transgene are silenced-a phenomenon known as position effect variegation (PEV). We show here that PEV can also occur when the transgene is non-centromeric and that the extent of variegation can be developmentally regulated. Furthermore, by overexpressing a mammalian homologue (M31) of Drosophila melanogaster heterochromatin protein 1 (HP1; refs 7,8) in transgenic mouse lines that exhibit PEV, it is possible to modify the proportion of cells that silence the transgene in a dose-dependent manner. Thus, we show M31 overexpression to have two contrasting effects which are dependent on chromosomal context: (i) it enhanced PEV in those lines with centromeric or pericentromeric transgene locations; and (ii) it suppressed PEV when the transgene was non-centromeric. Our results indicate that components or modifiers of heterochromatin may have a chromosomal-context-dependent role in gene silencing and activation decisions in mammals.     

TERRA and hnRNPA1 orchestrate an RPA-to-POT1 switch on telomeric single-stranded DNA

Maintenance of telomeres requires both DNA replication and telomere 'capping' by shelterin. These two processes use two single-stranded DNA (ssDNA)-binding proteins, replication protein A (RPA) and protection of telomeres 1 (POT1). Although RPA and POT1 each have a critical role at telomeres, how they function in concert is not clear. POT1 ablation leads to activation of the ataxia telangiectasia and Rad3-related (ATR) checkpoint kinase at telomeres, suggesting that POT1 antagonizes RPA binding to telomeric ssDNA. Unexpectedly, we found that purified POT1 and its functional partner TPP1 are unable to prevent RPA binding to telomeric ssDNA efficiently. In cell extracts, we identified a novel activity that specifically displaces RPA, but not POT1, from telomeric ssDNA. Using purified protein, here we show that the heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) recapitulates the RPA displacing activity. The RPA displacing activity is inhibited by the telomeric repeat-containing RNA (TERRA) in early S phase, but is then unleashed in late S phase when TERRA levels decline at telomeres. Interestingly, TERRA also promotes POT1 binding to telomeric ssDNA by removing hnRNPA1, suggesting that the re-accumulation of TERRA after S phase helps to complete the RPA-to-POT1 switch on telomeric ssDNA. Together, our data suggest that hnRNPA1, TERRA and POT1 act in concert to displace RPA from telomeric ssDNA after DNA replication, and promote telomere capping to preserve genomic integrity.     

Structural basis of PIP2 activation of the classical inward rectifier K+ channel Kir2.2

The regulation of ion channel activity by specific lipid molecules is widely recognized as an integral component of electrical signalling in cells. In particular, phosphatidylinositol 4,5-bisphosphate (PIP(2)), a minor yet dynamic phospholipid component of cell membranes, is known to regulate many different ion channels. PIP(2) is the primary agonist for classical inward rectifier (Kir2) channels, through which this lipid can regulate a cell's resting membrane potential. However, the molecular mechanism by which PIP(2) exerts its action is unknown. Here we present the X-ray crystal structure of a Kir2.2 channel in complex with a short-chain (dioctanoyl) derivative of PIP(2). We found that PIP(2) binds at an interface between the transmembrane domain (TMD) and the cytoplasmic domain (CTD). The PIP(2)-binding site consists of a conserved non-specific phospholipid-binding region in the TMD and a specific phosphatidylinositol-binding region in the CTD. On PIP(2) binding, a flexible expansion linker contracts to a compact helical structure, the CTD translates 6 ? and becomes tethered to the TMD and the inner helix gate begins to open. In contrast, the small anionic lipid dioctanoyl glycerol pyrophosphatidic acid (PPA) also binds to the non-specific TMD region, but not to the specific phosphatidylinositol region, and thus fails to engage the CTD or open the channel. Our results show how PIP(2) can control the resting membrane potential through a specific ion-channel-receptor-ligand interaction that brings about a large conformational change, analogous to neurotransmitter activation of ion channels at synapses.     

卫星导航设备时延精密标定方法与测试技术研究

分析和研究了卫星导航设备时延标定的方法,并针对卫星导航设备时延的特点,提出卫星导航设备时延应用统计量和稳定性来表征,并提出了基于射频采样和软件无线电的精密设备时延测量方法（RFSR）,能够准确地分析设备时延和设备时延的稳定性,通过仿真分析验证了方法正确性;通过建立试验验证系统,对可编程延迟线、电缆等设备的模拟测量,验证了该方法的正确性和优越性.     

Science,self improvement and the first industrial revolution

This study considers Newton's views on space and time with respect to some important ontologies of substance in his period. Specifically, it deals in a philosophico-historical manner with his conception of substance, attribute, existence, to actuality and necessity. I show how Newton links these “features” of things to his conception of God's existence with respect of infinite space and time. Moreover, I argue that his ontology of space and time cannot be understood without fully appreciating how it relates to the nature of Divine existence. In order to establish this, the ontology embodied in Newton's theory of predication is analysed, and shown to be different from the presuppositions of the ontological argument. From the historical point of view Gassendi's influence is stressed, via the mediation of Walter Charleton. Furthermore, Newton's thought on these matters is contrasted with Descartes's and Spinoza's. In point of fact, in his earliest notebook Newton recorded observations on Descartes's version of the ontological argument. Soon, however, he was to oppose the Cartesian conception of the actuality of Divine existence by means of arguments similar to those of Gassendi. Lastly, I suggest that the nature and extent of Henry More's influence on Newton's conception of how God relates to absolute space and time bears further examination.     

Francis Hauksbee's theory of electricity

Conclusion Historians of science have usually assumed that the science of electricity developed in the period prior to Franklin, or at least prior to Nollet, in what amounted to a theoretical vacuum. It has been my aim in this paper to demonstrate the falsity of that assumption. I have shown, I hope, that Hauksbee's important researches were guided throughout by strong theoretical considerations, and I have indicated that Dufay's even more important studies were guided by exactly the same considerations. Nor was their theory in any sense a stagnant one. As it was developed by Hauksbee, it could give a fairly adequate explanation of almost all the known electrical phenomena; it even enabled him to predict the outcome of experiments such as the one involving the rubbing of a globe while it was positioned near a second, exhausted, globe. With the discovery of so many new phenomena in the 1730's, the theory turned out to be no longer adequate, but it is not at all surprising that it was a few years before the full extent of its inadequacies was appreciated, nor is it surprising that a strong continuity is evident between it and the theory which eventually replaced it. In the meantime, the theory continued to serve a useful function by suggesting new lines of research to its adherents. The theory functioned, then, in the same way as any other scientific theory, and it deserves a more serious treatment than it has usually received. This paper, I hope, can serve as a beginning.     

Stage-specific sensitivity to p53 restoration during lung cancer progression

Tumorigenesis is a multistep process that results from the sequential accumulation of mutations in key oncogene and tumour suppressor pathways. Personalized cancer therapy that is based on targeting these underlying genetic abnormalities presupposes that sustained inactivation of tumour suppressors and activation of oncogenes is essential in advanced cancers. Mutations in the p53 tumour-suppressor pathway are common in human cancer and significant efforts towards pharmaceutical reactivation of defective p53 pathways are underway. Here we show that restoration of p53 in established murine lung tumours leads to significant but incomplete tumour cell loss specifically in malignant adenocarcinomas, but not in adenomas. We define amplification of MAPK signalling as a critical determinant of malignant progression and also a stimulator of Arf tumour-suppressor expression. The response to p53 restoration in this context is critically dependent on the expression of Arf. We propose that p53 not only limits malignant progression by suppressing the acquisition of alterations that lead to tumour progression, but also, in the context of p53 restoration, responds to increased oncogenic signalling to mediate tumour regression. Our observations also underscore that the p53 pathway is not engaged by low levels of oncogene activity that are sufficient for early stages of lung tumour development. These data suggest that restoration of pathways important in tumour progression, as opposed to initiation, may lead to incomplete tumour regression due to the stage-heterogeneity of tumour cell populations.     

Electron microscopic analysis of KvAP voltage-dependent K+ channels in an open conformation

Voltage-dependent ion channels serve as field-effect transistors by opening a gate in response to membrane voltage changes. The gate's response to voltage is mediated by voltage sensors, which are arginine-containing structures that must move with respect to the membrane electric field. We have analysed by electron microscopy a voltage-dependent K(+) channel from Aeropyrum pernix (KvAP). Fab fragments were attached to 'voltage sensor paddles' and identified in the electron microscopy map at 10.5 A resolution. The extracellular surface location of the Fab fragments in the map is consistent with the membrane-depolarized, open conformation of the channel in electrophysiological experiments. Comparison of the map with a crystal structure demonstrates that the voltage sensor paddles are 'up' (that is, near the channel's extracellular surface) and situated at the protein-lipid interface. This finding supports the hypothesis that in response to changes in voltage the sensors move at the protein-lipid interface rather than in a gating pore surrounded by protein.