Extension of life-span with superoxide dismutase/catalase mimetics

We tested the theory that reactive oxygen species cause aging. We augmented the natural antioxidant systems of Caenorhabditis elegans with small synthetic superoxide dismutase/catalase mimetics. Treatment of wild-type worms increased their mean life-span by a mean of 44 percent, and treatment of prematurely aging worms resulted in normalization of their life-span (a 67 percent increase). It appears that oxidative stress is a major determinant of life-span and that it can be counteracted by pharmacological intervention.     

Comparative genomics of the eukaryotes

A comparative analysis of the genomes of Drosophila melanogaster, Caenorhabditis elegans, and Saccharomyces cerevisiae-and the proteins they are predicted to encode-was undertaken in the context of cellular, developmental, and evolutionary processes. The nonredundant protein sets of flies and worms are similar in size and are only twice that of yeast, but different gene families are expanded in each genome, and the multidomain proteins and signaling pathways of the fly and worm are far more complex than those of yeast. The fly has orthologs to 177 of the 289 human disease genes examined and provides the foundation for rapid analysis of some of the basic processes involved in human disease.     

Augmented Wnt signaling in a mammalian model of accelerated aging

The contribution of stem and progenitor cell dysfunction and depletion in normal aging remains incompletely understood. We explored this concept in the Klotho mouse model of accelerated aging. Analysis of various tissues and organs from young Klotho mice revealed a decrease in stem cell number and an increase in progenitor cell senescence. Because klotho is a secreted protein, we postulated that klotho might interact with other soluble mediators of stem cells. We found that klotho bound to various Wnt family members. In a cell culture model, the Wnt-klotho interaction resulted in the suppression of Wnt biological activity. Tissues and organs from klotho-deficient animals showed evidence of increased Wnt signaling, and ectopic expression of klotho antagonized the activity of endogenous and exogenous Wnt. Both in vitro and in vivo, continuous Wnt exposure triggered accelerated cellular senescence. Thus, klotho appears to be a secreted Wnt antagonist and Wnt proteins have an unexpected role in mammalian aging.     

牛支原体蛋白质组学双向电泳技术的建立及初步分析

为了建立M.bovis蛋白质组学技术,试验采用M.bovis湖北分离株为实验材料,对样品处理、固相pH梯度胶条、上样量及染色等影响双向电泳图谱质量的关键因素与环节进行一系列的探索,2D Clean-up试剂盒处理的样品400μg在13 cm干胶条(pH 4~7)中进行双向凝胶电泳,考马斯亮蓝R350染色的方法,结果获得了能较好展示大部分蛋白点,分辨率及重复性均很好的双向电泳图谱,应用Image Master 2D Platinum version 6.0软件对图谱进行初步分析,不同重复图谱之间的平均匹配率达80.95%,随机取重复性好的20个蛋白点进行质谱分析,获得19蛋白质点的肽质量指纹图谱,鉴定成功率为95%。质谱鉴定结果与自建M.bovis蛋白库对比,蛋白点匹配率高于82分值限值(P<0.05)的不同蛋白有12个,初步分析这些蛋白多为酶类。说明该技术平台能有效用于M.bovis蛋白质组学的后续研究之中。     

PEG分级法检测绿竹叶片双向电泳中的低丰度蛋白

蛋白质组学研究的关键问题之一是双向电泳(2-DE)中低丰度蛋白的分析,主要是因为相关高丰度蛋白的存在,导致IPG胶条无法吸胀低丰度蛋白,使得低丰度蛋白在2-DE中很难被检测出来。利用梯度浓度的PEG4000沉淀方法来富集绿竹叶片中不同丰度的蛋白质,从而使得低丰度蛋白在凝胶中显现出来。在PEG分级法和传统的全蛋白提取法的2-DE比较中,发现采用PEG分级沉淀法提取的蛋白质的数量以及类别明显增加,高丰度蛋白主要富集在8%和16%的PEG浓度组分中,使得其他组分中的低丰度蛋白与高丰度蛋白组分分别进行2-DE分析。经过对图像和数据的比较、分析,PEG分级法得到的5个组分蛋白点总数超过了1032个,大约是传统蛋白提取方法制备蛋白样品的3倍。     

应用双向电泳技术研究苹果实生树童期和成年期叶片蛋白质的变化

 为了研究苹果实生树阶段转变过程中蛋白质的定性和定量变化规律,利用双向电泳（two dimensional electrophoresis, 2 DE）技术对苹果实生树童期和成年期叶片的蛋白质组分进行了研究。结果表明,童期和成年期的2 DE图谱上,有23个蛋白质点在表达上存在明显的质和量的变化。其中,6个蛋白质点为童期组织所特有,3个蛋白质点只能在成年期叶片中检测到。此外,有14个蛋白质点在表达水平上存在显著差异,即在成年期叶片中有11个蛋白质点的含量显著高于童期;有3个蛋白质点的含量明显低于童期叶片。因此认为检测到的23个差异蛋白质点可能与阶段转变有关。     

植物线粒体和质体双定位蛋白质的带电特性分析

蛋白质亚细胞定位分析是揭示蛋白质功能的关键步骤。1个蛋白质分子能被定位到2个亚细胞位置,这一现象被称为蛋白质的"双定位"。本研究首先从Uniprot、MitoP2、MGI、TAIR、DBMLoc等蛋白质数据库及已发表文献中收集双定位于线粒体与质体的植物蛋白质数据,共获得703个双定位蛋白质,组成测试数据集。再从Uniprot数据库中选取唯一定位于线粒体的829个和唯一定位于质体的6 376个植物蛋白质,组成参照数据集,分析双定位于线粒体与质体的植物蛋白质的带电特征。结果表明,与单定位于线粒体或质体的植物蛋白质相比,双定位线粒体与质体的植物蛋白质具有更低的净电荷量;此外,双定位蛋白质电荷分布较为集中对称,线粒体蛋白质次之,质体蛋白质最为分散。本文研究结果将为揭示植物蛋白质双定位的分子机制奠定理论基础。     

Molecular biology. Unwinding RNA silencing

Eukaryotic cells have developed an elegant system called RNA silencing for getting rid of foreign RNAs whether they be of viral, retrotransposon, or transgene origin. In his Perspective, Baulcombe examines new findings (Wu-Scharf et al.) showing that in a green alga the gene responsible for RNA silencing encodes an RNA helicase (related to proteins in worms and other organisms) that is required for regulation of gene expression at the RNA level.     

两种花吊丝竹叶片蛋白提取方法的2-DE比较

以2年生花吊丝竹的扦插苗为研究材料,对2种方法(TCA/酚提结合法和PEG法)提取的蛋白质的2-DE差异进行研究.结果表明:PEG法的5个组分的条带差异明显,全蛋白得到了有效分离,在F3中富集到了2条高丰度蛋白带;2种方法的2-DE图谱的均一性分析发现,PEG法中F3富集了高丰度蛋白,提高了2-DE中蛋白质的检测率和分辨率,PEG法中F1-F5共检测的蛋白总数超过了1700个,但是相邻组存在一定的重叠率,而TCA丙酮/酚提结合法只检测到约571个蛋白点,5个组与NF平均有439个蛋白点匹配,达到77.9%以上;质谱鉴定了5种差异蛋白,生物信息学分析发现这5种蛋白是糖酵解、糖异生和ATP合成的重要辅酶.2种方法的比较发现,PEG法能够将花吊丝竹叶片全蛋白中高丰度蛋白单一富集在16%的组分中,从而显著提高了2-DE的分辨率,再通过质谱技术能够检测到更多的影响生物体生长发育的低丰度蛋白,所以PEG法是一种切实可行的且较为理想的蛋白质提取方法.     

Mutations that increase the life span of C. elegans inhibit tumor growth

Mutations in gld-1 cause lethal germline tumors in the nematode Caenorhabditis elegans. We find that a wide variety of mutations that extend C. elegans' life span confer resistance to these tumors. The long life spans of daf-2/insulin-receptor mutants were not shortened at all by gld-1 mutations; we attribute this finding to decreased cell division and increased DAF-16/p53-dependent apoptosis within the tumors. Mutations that increase life span by restricting food intake or inhibiting respiration did not affect apoptosis but reduced tumor cell division. Unexpectedly, none of these longevity mutations affected mitosis in normal germlines; this finding suggests that cellular changes that lead to longevity preferentially antagonize tumor cell growth.     

花生种子人工老化过程中发芽率和蛋白质的变化

利用双向ＳＤＳ－ＰＡＧＥ分析了粤油７９和油７３花生种子人工老化过程中发芽率和蛋白质的变化，结果表明，种子老化８ｄ后，种胚内出现了一种蛋白质或多肽（ｐ１６．２，ＭＷ１０ＫＤ），并且该蛋白质或多肽随种子老化程度的加深其含量逐渐增加，而对照不存在这种物质，表明随着种子活力的丧失，蛋白质的含量增加。     

斯氏线虫对两种地老虎的侵染效果

１９９１￣１９９３年室内试验结果证明：芜菁夜蛾线虫的３个品系Ｂｅｉｊｉｎｇ、Ｍｅｘｃｉａｎ、Ｏｔｉｏ对八字地老虎和小地老虎两种地下害虫的幼虫有很高的侵染力．用Ｂｅｉｊｉｎｇ品系线虫处理八字地老虎３龄幼虫，幼虫死亡率随线虫剂量而增加，当剂量为８０条线虫／八字地老虎幼虫时．死亡率可达９３％以上．每头３龄小地老虎幼虫用８０条该线虫处理的死亡率为８０％，用同一剂量的Ｍｅｘｉｃａｎ品系，八字地老虎和小地老虎的死亡率分别为８６．６％和８５．１％．Ｏｔｉｏ品系对两种幼虫的致死率达７０％￣８３．３％．ＮＣ３２品系对八字老虎毒力较差．即使剂量增至１６００条线虫／虫，八字地老虎的死亡率仅为４０％，表明不同品系线虫对害虫毒力差异较大．     

Substitution of the thioredoxin system for glutathione reductase in Drosophila melanogaster

The disulfide reducing enzymes glutathione reductase and thioredoxin reductase are highly conserved among bacteria, fungi, worms, and mammals. These proteins maintain intracellular redox homeostasis to protect the organism from oxidative damage. Here we demonstrate the absence of glutathione reductase in Drosophila melanogaster, identify a new type of thioredoxin reductase, and provide evidence that a thioredoxin system supports GSSG reduction. Our data suggest that antioxidant defense in Drosophila, and probably in related insects, differs fundamentally from that in other organisms.