Role of adenine nucleotide translocator 1 in mtDNA maintenance

Autosomal dominant progressive external ophthalmoplegia is a rare human disease that shows a Mendelian inheritance pattern, but is characterized by large-scale mitochondrial DNA (mtDNA) deletions. We have identified two heterozygous missense mutations in the nuclear gene encoding the heart/skeletal muscle isoform of the adenine nucleotide translocator (ANT1) in five families and one sporadic patient. The familial mutation substitutes a proline for a highly conserved alanine at position 114 in the ANT1 protein. The analogous mutation in yeast caused a respiratory defect. These results indicate that ANT has a role in mtDNA maintenance and that a mitochondrial disease can be caused by a dominant mechanism.     

Discovery of a major D-loop replication origin reveals two modes of human mtDNA synthesis

Mammalian mitochondrial DNA (mtDNA) replication has long been considered to occur by asymmetric synthesis of the two strands, starting at the multiple origins of the strand-displacement loop (D-loop). We report the discovery of a major replication origin at position 57 in the D-loop of several human cell lines (HeLa, A549, and 143B.TK-) and immortalized lymphocytes. The nascent chains starting at this origin, in contrast to those initiated at the previously described origins, do not terminate prematurely at the 3' end of the D-loop but proceed well beyond this control point, behaving as "true" replicating strands. This origin is mainly responsible for mtDNA maintenance under steady-state conditions, whereas mtDNA synthesis from the formerly identified D-loop origins may be more important for recovery after mtDNA depletion and for accelerating mtDNA replication in response to physiological demands.     

Comment on "Population size does not influence mitochondrial genetic diversity in animals"

Bazin et al. (Reports, 28 April, 2006, p. 570) found no relationship between mitochondrial DNA (mtDNA) diversity and population size when comparing across large groups of animals. We show empirically that species with smaller populations, as represented by eutherian mammals, exhibit a positive correlation between mtDNA and allozyme variation, suggesting that mtDNA diversity may correlate with population size in these animals.     

Facile detection of mitochondrial DNA mutations in tumors and bodily fluids

Examination of human bladder, head and neck, and lung primary tumors revealed a high frequency of mitochondrial DNA (mtDNA) mutations. The majority of these somatic mutations were homoplasmic in nature, indicating that the mutant mtDNA became dominant in tumor cells. The mutated mtDNA was readily detectable in paired bodily fluids from each type of cancer and was 19 to 220 times as abundant as mutated nuclear p53 DNA. By virtue of their clonal nature and high copy number, mitochondrial mutations may provide a powerful molecular marker for noninvasive detection of cancer.     

道寒杂交公羊育肥期能量需要量研究

【目的】确定道寒杂交公羊育肥期维持和生长的净能与代谢能需要量参数,为肉羊的科学饲养提供理论依据。【方法】本试验选用34只年龄相近（6月龄）、平均体重(34.54±0.40)kg的无角道赛特和小尾寒羊杂交公羔为试验羊。其中25只用于比较屠宰试验,随机分为自由采食组（15只）、60%自由采食组（5只）、40%自由采食组（5只）,其中自由采食组又随机分为3组,每组5只。在正式试验开始后的第1天、第19 天（平均体重43 kg）、第63 天（平均体重50 kg）分别从自由采食第一组、第二组、第三组中选择体重接近的4只试验羊进行屠宰,同时,从60%自由采食组、40%自由采食组中分别选择体重接近的4只试验羊与自由采食第三组一起屠宰。另外9只用于消化代谢试验,同样按上述饲喂水平随机分为3组,采用全收粪尿法收集粪、尿,利用推荐公式对试验羊的甲烷能进行预测。【结果】道寒杂交公羊育肥期的维持净能需要量为286.88 kJ•kg-1 EBW0.75（269.09 kJ•kg-1 BW0.75）,维持代谢能需要量为423.89 kJ•kg-1 EBW0.75（397.60 kJ•kg-1 BW0.75）。日增重100—350 g时生长净能和生长代谢能需要量分别为1.45—6.20 MJ•d-1、3.21—13.72 MJ•d-1。代谢能维持利用效率为0.68,代谢能生长利用效率为0.45。【结论】道寒杂交公羊育肥期能量代谢参数值与NRC（2007）推荐的能量需要量相比有一定差异。     

Structure of nerve growth factor complexed with the shared neurotrophin receptor p75

Neurotrophins are secreted growth factors critical for the development and maintenance of the vertebrate nervous system. Neurotrophins activate two types of cell surface receptors, the Trk receptor tyrosine kinases and the shared p75 neurotrophin receptor. We have determined the 2.4 A crystal structure of the prototypic neurotrophin, nerve growth factor (NGF), complexed with the extracellular domain of p75. Surprisingly, the complex is composed of an NGF homodimer asymmetrically bound to a single p75. p75 binds along the homodimeric interface of NGF, which disables NGF's symmetry-related second p75 binding site through an allosteric conformational change. Thus, neurotrophin signaling through p75 may occur by disassembly of p75 dimers and assembly of asymmetric 2:1 neurotrophin/p75 complexes, which could potentially engage a Trk receptor to form a trimolecular signaling complex.     

20-35 kg杜寒杂交公羔羊能量需要参数

[目的]旨在确定20-35 kg杜寒杂交F1代公羔羊维持和生长的代谢能与净能需要量,为杜寒杂交羔羊的科学饲养提供参考依据.[方法]本试验采用完全随机区组试验设计,正试期66 d.筛选基本同龄的杜寒杂交公羔羊50只,其中35只用于比较屠宰试验,15只用于消化代谢试验.当用于比较屠宰试验的35只羊体重为(20.26±1.29)kg时,随机抽取7只进行屠宰用以估测试验羊初始体组成;剩余的28只羊分为自由采食组(AL,14只)、70％自由采食组(IR70,7只)和40％自由采食组(IR40,7只)3个饲喂水平组.当自由采食组的14只羊体重达到(28.54±2.29) kg时,随机抽取7只羊进行屠宰;将剩余的21只羊按上述3个饲喂水平饲喂,每个饲喂水平包括7只羊.当自由采食组羊只平均体重达到35 kg时,按屠宰规程进行屠宰.在消化代谢试验中,将体重为(32.38±2.23) kg的15只杜寒杂交公羔羊随机分为3个处理组(每个处理组5只羊),试验日粮及饲喂水平同比较屠宰试验,采用全收粪尿法收集粪、尿,采用开放式呼吸测热系统测定24 h甲烷产生量、CO2排放量和耗氧量.[结果]杜寒杂交公羔羊维持净能和维持代谢能需要量分别为250.61和374.21 kJ·kg-1SBW0.75,代谢能维持利用效率为0.67;在20-35 kg体重阶段,日增重分别为100、200、300和350 g·d-1生长净能和生长代谢能的需要量变化范围在1.10-5.04 MJ.d-1和2.63-12.03 MJ·d-1,代谢能的生长利用效率为0.419.[结论]20-35 kg杜寒杂交F1代公羔羊能量代谢参数(包括维持净能、维持代谢能、生长净能和生长代谢能)参数值略低于NRC和AFRC肉羊能量需要量的推荐标准.     

p53 regulates mitochondrial respiration

The energy that sustains cancer cells is derived preferentially from glycolysis. This metabolic change, the Warburg effect, was one of the first alterations in cancer cells recognized as conferring a survival advantage. Here, we show that p53, one of the most frequently mutated genes in cancers, modulates the balance between the utilization of respiratory and glycolytic pathways. We identify Synthesis of Cytochrome c Oxidase 2 (SCO2) as the downstream mediator of this effect in mice and human cancer cell lines. SCO2 is critical for regulating the cytochrome c oxidase (COX) complex, the major site of oxygen utilization in the eukaryotic cell. Disruption of the SCO2 gene in human cancer cells with wild-type p53 recapitulated the metabolic switch toward glycolysis that is exhibited by p53-deficient cells. That SCO2 couples p53 to mitochondrial respiration provides a possible explanation for the Warburg effect and offers new clues as to how p53 might affect aging and metabolism.     

烟粉虱复合种不同地理种群的遗传分化

烟粉虱Bemisia tabaci(Gennadius) 是由具有遗传分化的不同地理种群组成的复合种。本文介绍了烟粉虱不同地理种群遗传分化的最新研究进展，并在世界各国对烟粉虱核糖体ITS1(rDNA ITS1)、线粒体COI（mtDNA COI）基因大量测序的基础上，进一步分析了烟粉虱不同地理种群的遗传分化。根据mtDNA COI和rDNA ITS1基因序列分析的结果，烟粉虱不同地理种群可分为5组，即亚洲组（Asia group）、美洲组（America group）、非洲组（Africa group）、澳洲组(Australia group)、B型/地中海/中东/北非/Ms型组(Biotype B/Mediterranean/Middle Eastern/Northern Africa/Biotype Ms group)；此外，还包括3个没有特定组的种群，即乌干达（Uganda）、科特迪瓦（Ivory Coast和台湾（Taiwan）种群。地理隔离可能是造成烟粉虱不同地理种群遗传分化的最重要驱动力。许多具有入侵性或生物学优势的烟粉虱种群随着人类活动而传入新的地区，造成了严重的经济损失。有必要加强烟粉虱生物型的监测，遏制已入侵烟粉虱种群的蔓延，防止新的具有潜在危险性的烟粉虱种群传入中国。     

Noncytotoxic lytic granule-mediated CD8+ T cell inhibition of HSV-1 reactivation from neuronal latency

Reactivation of herpes simplex virus type 1 (HSV-1) from neuronal latency is a common and potentially devastating cause of disease worldwide. CD8+ T cells can completely inhibit HSV reactivation in mice, with interferon-gamma affording a portion of this protection. We found that CD8+ T cell lytic granules are also required for the maintenance of neuronal latency both in vivo and in ex vivo ganglia cultures and that their directed release to the junction with neurons in latently infected ganglia did not induce neuronal apoptosis. Here, we describe a nonlethal mechanism of viral inactivation in which the lytic granule component, granzyme B, degrades the HSV-1 immediate early protein, ICP4, which is essential for further viral gene expression.     

Ancient DNA from the first European farmers in 7500-year-old Neolithic sites

The ancestry of modern Europeans is a subject of debate among geneticists, archaeologists, and anthropologists. A crucial question is the extent to which Europeans are descended from the first European farmers in the Neolithic Age 7500 years ago or from Paleolithic hunter-gatherers who were present in Europe since 40,000 years ago. Here we present an analysis of ancient DNA from early European farmers. We successfully extracted and sequenced intact stretches of maternally inherited mitochondrial DNA (mtDNA) from 24 out of 57 Neolithic skeletons from various locations in Germany, Austria, and Hungary. We found that 25% of the Neolithic farmers had one characteristic mtDNA type and that this type formerly was widespread among Neolithic farmers in Central Europe. Europeans today have a 150-times lower frequency (0.2%) of this mtDNA type, revealing that these first Neolithic farmers did not have a strong genetic influence on modern European female lineages. Our finding lends weight to a proposed Paleolithic ancestry for modern Europeans.     

ROS-generating mitochondrial DNA mutations can regulate tumor cell metastasis

Mutations in mitochondrial DNA (mtDNA) occur at high frequency in human tumors, but whether these mutations alter tumor cell behavior has been unclear. We used cytoplasmic hybrid (cybrid) technology to replace the endogenous mtDNA in a mouse tumor cell line that was poorly metastatic with mtDNA from a cell line that was highly metastatic, and vice versa. Using assays of metastasis in mice, we found that the recipient tumor cells acquired the metastatic potential of the transferred mtDNA. The mtDNA conferring high metastatic potential contained G13997A and 13885insC mutations in the gene encoding NADH (reduced form of nicotinamide adenine dinucleotide) dehydrogenase subunit 6 (ND6). These mutations produced a deficiency in respiratory complex I activity and were associated with overproduction of reactive oxygen species (ROS). Pretreatment of the highly metastatic tumor cells with ROS scavengers suppressed their metastatic potential in mice. These results indicate that mtDNA mutations can contribute to tumor progression by enhancing the metastatic potential of tumor cells.     

3个罗非鱼品系的线粒体DNA遗传多态性研究

应用mtDNA标记技术对3个罗非鱼品种(系)(吉富品系尼罗罗非鱼、美国品系尼罗罗非鱼、埃及品系尼罗罗非鱼)的DNA多态性进行研究.研究结果如下:(1)测定3个品种的罗非鱼mtDNA Cytb基因的一段长为435 bp的序列.发现2个碱基发生转换,1个氨基酸被替换;(2)将3个品种的罗非鱼mtDNA Cytb基因部分序列与马氏蛰丽鱼、迪氏康尼丽鱼、洛氏帚齿非鲫、加利略帚齿非鲫、彩虹鲷、莫桑比克共9个个体的mtDNA Cytb基因进行同源序列分析,系统关系可表示为:[(尼罗罗非鱼,莫桑比克罗非鱼),马氏蛰丽鱼],这与传统的分类学结果一致.     

Comment on "Whole-genome shotgun sequencing of mitochondria from ancient hair shafts"

Gilbert et al. (Reports, 28 September 2007, p. 1927) reported that "hair shafts surpass comparably stored bone as an aDNA source [...] in regard to preservation and concentration of mtDNA." When experimental parameters are carefully controlled for, including adequate sampling, quantitative polymerase chain reaction analysis, and modeling the decay of DNA, the general importance of this claim is not supported.     

Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex

The complex containing the Mre11, Rad50, and Nbs1 proteins (MRN) is essential for the cellular response to DNA double-strand breaks, integrating DNA repair with the activation of checkpoint signaling through the protein kinase ATM (ataxia telangiectasia mutated). We demonstrate that MRN stimulates the kinase activity of ATM in vitro toward its substrates p53, Chk2, and histone H2AX. MRN makes multiple contacts with ATM and appears to stimulate ATM activity by facilitating the stable binding of substrates. Phosphorylation of Nbs1 is critical for MRN stimulation of ATM activity toward Chk2, but not p53. Kinase-deficient ATM inhibits wild-type ATM phosphorylation of Chk2, consistent with the dominant-negative effect of kinase-deficient ATM in vivo.     

Mitochondrial DNA mutations, oxidative stress, and apoptosis in mammalian aging

Mutations in mitochondrial DNA (mtDNA) accumulate in tissues of mammalian species and have been hypothesized to contribute to aging. We show that mice expressing a proofreading-deficient version of the mitochondrial DNA polymerase g (POLG) accumulate mtDNA mutations and display features of accelerated aging. Accumulation of mtDNA mutations was not associated with increased markers of oxidative stress or a defect in cellular proliferation, but was correlated with the induction of apoptotic markers, particularly in tissues characterized by rapid cellular turnover. The levels of apoptotic markers were also found to increase during aging in normal mice. Thus, accumulation of mtDNA mutations that promote apoptosis may be a central mechanism driving mammalian aging.