A large-scale RNAi screen in human cells identifies new components of the p53 pathway

RNA interference (RNAi) is a powerful new tool with which to perform loss-of-function genetic screens in lower organisms and can greatly facilitate the identification of components of cellular signalling pathways. In mammalian cells, such screens have been hampered by a lack of suitable tools that can be used on a large scale. We and others have recently developed expression vectors to direct the synthesis of short hairpin RNAs (shRNAs) that act as short interfering RNA (siRNA)-like molecules to stably suppress gene expression. Here we report the construction of a set of retroviral vectors encoding 23,742 distinct shRNAs, which target 7,914 different human genes for suppression. We use this RNAi library in human cells to identify one known and five new modulators of p53-dependent proliferation arrest. Suppression of these genes confers resistance to both p53-dependent and p19ARF-dependent proliferation arrest, and abolishes a DNA-damage-induced G1 cell-cycle arrest. Furthermore, we describe siRNA bar-code screens to rapidly identify individual siRNA vectors associated with a specific phenotype. These new tools will greatly facilitate large-scale loss-of-function genetic screens in mammalian cells.     

关于停时的一个注记

对任意停时T,定义A(T)={停时S:S≤T,在{T>0}上S相似文献   

Heat activation of TRPM5 underlies thermal sensitivity of sweet taste

TRPM5, a cation channel of the TRP superfamily, is highly expressed in taste buds of the tongue, where it has a key role in the perception of sweet, umami and bitter tastes. Activation of TRPM5 occurs downstream of the activation of G-protein-coupled taste receptors and is proposed to generate a depolarizing potential in the taste receptor cells. Factors that modulate TRPM5 activity are therefore expected to influence taste. Here we show that TRPM5 is a highly temperature-sensitive, heat-activated channel: inward TRPM5 currents increase steeply at temperatures between 15 and 35 degrees C. TRPM4, a close homologue of TRPM5, shows similar temperature sensitivity. Heat activation is due to a temperature-dependent shift of the activation curve, in analogy to other thermosensitive TRP channels. Moreover, we show that increasing temperature between 15 and 35 degrees C markedly enhances the gustatory nerve response to sweet compounds in wild-type but not in Trpm5 knockout mice. The strong temperature sensitivity of TRPM5 may underlie known effects of temperature on perceived taste in humans, including enhanced sweetness perception at high temperatures and 'thermal taste', the phenomenon whereby heating or cooling of the tongue evoke sensations of taste in the absence of tastants.     

A new hominid from the Upper Miocene of Chad,Central Africa

The search for the earliest fossil evidence of the human lineage has been concentrated in East Africa. Here we report the discovery of six hominid specimens from Chad, central Africa, 2,500 km from the East African Rift Valley. The fossils include a nearly complete cranium and fragmentary lower jaws. The associated fauna suggest the fossils are between 6 and 7 million years old. The fossils display a unique mosaic of primitive and derived characters, and constitute a new genus and species of hominid. The distance from the Rift Valley, and the great antiquity of the fossils, suggest that the earliest members of the hominid clade were more widely distributed than has been thought, and that the divergence between the human and chimpanzee lineages was earlier than indicated by most molecular studies.     

水分和热胁迫对苗木针叶可溶性糖含量的影响

以杉木、马尾松、花旗松和北美乔柏为对象，研究了在水分和热胁迫下，苗木生长期间针叶内可溶性糖含量的变化。结果表明：(1)水分和热胁迫处理对杉木针叶果糖含量的影响不明显，但对杉木针叶葡萄糖和蔗糖的含量却有显影响。水分胁迫和热胁迫的交互作用对杉苗针叶的蔗糖含量也有显影响。随着水分胁迫程度的加深，葡萄糖含量提高了11．80％，蔗糖含量提高幅度达318．86％。随着热胁迫程度的加深，葡萄糖含量提高了16．54％，蔗糖含量提高幅度为169．83％；(2)水分胁迫处理对马尾松苗木针叶葡萄糖和蔗糖含量的影响显，而热胁迫则对葡萄糖和果糖含量影响显。水分和热胁迫处理对马尾松苗木针叶3种可溶性糖含量的影响均呈上升趋势，其中水分胁迫条件下葡萄糖含量提高了45．92％，果糖含量提高了34．01％，蔗糖含量提高了138．32％。热胁迫条件下葡萄糖含量提高了36．07％，果糖含量提高了28．01％，蔗糖含量提高了22．82％；(3)水分和热胁迫尽管对花旗松苗木针叶的3种可溶性糖含量均有较显影响，但随着水分胁迫程度的加深，3种可溶性糖含量均未表现出明显的上升趋势。只有随着热胁迫程度的加深，花旗松苗木针叶的3种可溶性糖含量才呈现出上升趋势；(4)水分和热胁迫对北美乔柏苗木针叶可溶性糖含量均有显影响。其中随着水分胁迫程度的加深，葡萄糖含量提高了89．46％，果糖含量提高了90．32％，蔗糖含量提高了71.68％。随着热胁迫程度的加深，葡萄糖含量提高了21.18％，果糖含量提高了45．97％，蔗糖含量提高了107．67％。水分胁迫和热胁迫的交互作用对北美乔柏苗木针叶的蔗糖含量有显影响。     

可编程逻辑互连网络的设计

半导体技术的迅速进步要求数字系统设计过程的同步提升。例如英特尔4004处理器这样的早期集成电路完全是手工设计的，其中包括了布局工艺图。对于在1971年利用100m技术工艺过程构建的2300个晶体管晶件而言，这是一个合理的努力。与之形成对照的是2002年7月公布的最新的英特尔奔腾2微处理器。它包含了2．2亿个晶体管，使用了0．180m工艺过程，设计这样一个大型器件要求几百个工程师依靠高级的CAD工具来处理其复杂性。在过去的5至10年内，每一个新的英特尔处理器都是设计的顶峰，为其他器件的跟进设置了标准。随着当前制造技术达到了深一亚微层次，为创建一个可以工作的设计所要求的低层次设计努力就越来越多。同时可编程逻辑器件的功能已经提升。目前的现场可编程门阵列（FPGA）可以实现整个数字系统，它们对于系统层次设计者的吸引力在不断地增加。但是在许多情况中，它们的性能是不够的，或者它们没有包含足够的内存，或者缺乏关键的知识产权核心。因此存在着不断增加的需求，要把可编程逻辑技术与客户化的亚微VLSI技术相结合。如果说在可编程逻辑中存在着一个没有被完全理解的具体方面的话，那就是互连。它占用了最大的面积而且要为大多数的延时负责。可编程逻辑器件的90％是布局，10％是逻辑。本书把焦点放在了90％上面，即可编程布局上。它介绍了互连设计的最新研究成果，重点是互连结构自动生成的知识与工具。     

The sirtuin SIRT6 regulates lifespan in male mice

The significant increase in human lifespan during the past century confronts us with great medical challenges. To meet these challenges, the mechanisms that determine healthy ageing must be understood and controlled. Sirtuins are highly conserved deacetylases that have been shown to regulate lifespan in yeast, nematodes and fruitflies. However, the role of sirtuins in regulating worm and fly lifespan has recently become controversial. Moreover, the role of the seven mammalian sirtuins, SIRT1 to SIRT7 (homologues of the yeast sirtuin Sir2), in regulating lifespan is unclear. Here we show that male, but not female, transgenic mice overexpressing Sirt6 (ref. 4) have a significantly longer lifespan than wild-type mice. Gene expression analysis revealed significant differences between male Sirt6-transgenic mice and male wild-type mice: transgenic males displayed lower serum levels of insulin-like growth factor 1 (IGF1), higher levels of IGF-binding protein 1 and altered phosphorylation levels of major components of IGF1 signalling, a key pathway in the regulation of lifespan. This study shows the regulation of mammalian lifespan by a sirtuin family member and has important therapeutic implications for age-related diseases.     

Mutations in NALP7 cause recurrent hydatidiform moles and reproductive wastage in humans

Hydatidiform mole (HM) is an abnormal human pregnancy with no embryo and cystic degeneration of placental villi. We report five mutations in the maternal gene NALP7 in individuals with familial and recurrent HMs. NALP7 is a member of the CATERPILLER protein family involved in inflammation and apoptosis. NALP7 is the first maternal effect gene identified in humans and is also responsible for recurrent spontaneous abortions, stillbirths and intrauterine growth retardation.