The Trypanosoma cruzi proteome

To complement the sequencing of the three kinetoplastid genomes reported in this issue, we have undertaken a whole-organism, proteomic analysis of the four life-cycle stages of Trypanosoma cruzi. Peptides mapping to 2784 proteins in 1168 protein groups from the annotated T. cruzi genome were identified across the four life-cycle stages. Protein products were identified from >1000 genes annotated as "hypothetical" in the sequenced genome, including members of a newly defined gene family annotated as mucin-associated surface proteins. The four parasite stages appear to use distinct energy sources, including histidine for stages present in the insect vectors and fatty acids by intracellular amastigotes.     

Destruction of Trypanosoma cruzi by Natural killer cells

Mice infected with trypanosoma cruzi or stimulated with poly(inosinic.cytodylic acid) were found to possess splenic and peritoneal exudate cells with enhanced cytotoxic activity against epimastigote and trypomastigote forms of Trypanosoma cruzi. By use of specific alloantiserums it was determined that the effector cells responsible for this cytotoxic activity were typical natural killer cells.     

Inhibitory action of gossypol on enzymes and growth of Trypanosoma cruzi

Gossypol, a phenolic compound isolated from the cotton plant, is a powerful inhibitor of nicotinamide adenine dinucleotide-linked enzymes (alpha-hydroxyacid dehydrogenase and malate dehydrogenase) of Trypanosoma cruzi, the parasite that causes Chagas' disease. Parasites at the epimastigote stage that were incubated for 5 minutes with 100 micromolar gossypol were completely immobilized. Concentrations of gossypol as low as 0.01 micromolar markedly reduced the growth rate of T. cruzi in culture.     

The genome sequence of Trypanosoma cruzi, etiologic agent of Chagas disease

Whole-genome sequencing of the protozoan pathogen Trypanosoma cruzi revealed that the diploid genome contains a predicted 22,570 proteins encoded by genes, of which 12,570 represent allelic pairs. Over 50% of the genome consists of repeated sequences, such as retrotransposons and genes for large families of surface molecules, which include trans-sialidases, mucins, gp63s, and a large novel family (>1300 copies) of mucin-associated surface protein (MASP) genes. Analyses of the T. cruzi, T. brucei, and Leishmania major (Tritryp) genomes imply differences from other eukaryotes in DNA repair and initiation of replication and reflect their unusual mitochondrial DNA. Although the Tritryp lack several classes of signaling molecules, their kinomes contain a large and diverse set of protein kinases and phosphatases; their size and diversity imply previously unknown interactions and regulatory processes, which may be targets for intervention.     

Trypanosoma cruzi infection inhibited by peptides modeled from a fibronectin cell attachment domain

The mechanism by which Trypanosoma cruzi, the protozoan parasite that causes Chagas' disease, becomes attached to mammalian cells is not well understood. Fibronectin is thought to participate in the attachment, and in this study the region of fibronectin that interacts with the surface receptors of T. cruzi trypomastigotes was investigated by testing the binding of the amino acid sequence Arg-Gly-Asp-Ser, corresponding to the cell attachment site of fibronectin to T. cruzi trypomastigotes. Peptides with the sequence Arg-Gly-Asp-Ser, but not Arg-Phe-Asp-Ser, Arg-Phe-Asp-Ser-Ala-Ala-Arg-Phe-Asp, Ser-Lys-Pro, Glu-Ser-Gly, or Ala-Lys-Thr-Lys-Pro, bound to the parasite surface and inhibited cell invasion by the pathogen. Monoclonal antibodies to the cell attachment domain of fibronectin also inhibited cell infection by the parasite. The immunization of BALB/c mice with tetanus toxoid-conjugated peptide induced a significant protection against T. cruzi. The data support the notion that the sequence Arg-Gly-Asp-Ser of cell surface fibronectin acts as a recognition site for attachment of the parasites.     

Pars intermedia: unitary electrical activity regulated by light

In the pars intermedia of frogs in the dark two types of spontaneously firing neuronal units have been found; one can be inhibited by and the other is indifferent to increases in illumination. The receptor for the light-inhibited units appears to be the pineal organ. Transection experiments indicate that the axons to the two kinds of units in the pars intermedia are separately grouped in the floor.     

叶绿体硫氧还蛋白调控谷氨酸-1-半醛氨基转移酶氧化还原状态的研究

5-氨基乙酰丙酸(5-aminolevulinic acid,ALA)是四吡咯化合物合成的前体分子,它以谷氨酰-tRNA为底物经过一个两步反应形成,分别由谷氨酰-tRNA还原酶(glutamyl-tRNA reductase,GLTR)与谷氨酸-1-半醛氨基转移酶(Glutamate-1-semialdehyde aminotransferase,GSAAT)催化。GSAAT作为调控ALA合成的酶之一,对四吡咯化合物的合成起着至关重要的作用。硫氧还蛋白是一类小分子量的氧化还原蛋白(大约12kD)。它们介导其靶蛋白中半胱氨酸的二硫键与巯基之间的转变从而调控其靶蛋白的功能和稳定性。酵母双杂交试验结果显示,GSAAT与叶绿体硫氧还蛋白F(Thioredoxin-F)发生相互作用。利用VIGS技术沉默豌豆叶片中的叶绿体硫氧还蛋白基因,借助Western Blotting分析体内GSAAT的氧化还原状态,发现叶绿体硫氧还蛋白基因的沉默影响了GSAAT的氧化还原状态。     

A haptoglobin-hemoglobin receptor conveys innate immunity to Trypanosoma brucei in humans

The protozoan parasite Trypanosoma brucei is lysed by apolipoprotein L-I, a component of human high-density lipoprotein (HDL) particles that are also characterized by the presence of haptoglobin-related protein. We report that this process is mediated by a parasite glycoprotein receptor, which binds the haptoglobin-hemoglobin complex with high affinity for the uptake and incorporation of heme into intracellular hemoproteins. In mice, this receptor was required for optimal parasite growth and the resistance of parasites to the oxidative burst by host macrophages. In humans, the trypanosome receptor also recognized the complex between hemoglobin and haptoglobin-related protein, which explains its ability to capture trypanolytic HDLs. Thus, in humans the presence of haptoglobin-related protein has diverted the function of the trypanosome haptoglobin-hemoglobin receptor to elicit innate host immunity against the parasite.     

A photon turnstile dynamically regulated by one atom

Beyond traditional nonlinear optics with large numbers of atoms and photons, qualitatively new phenomena arise in a quantum regime of strong interactions between single atoms and photons. By using a microscopic optical resonator, we achieved such interactions and demonstrated a robust, efficient mechanism for the regulated transport of photons one by one. With critical coupling of the input light, a single atom within the resonator dynamically controls the cavity output conditioned on the photon number at the input, thereby functioning as a photon turnstile. We verified the transformation from a Poissonian to a sub-Poissonian photon stream by photon counting measurements of the input and output fields. The results have applications in quantum information science, including for controlled interactions of single light quanta and for scalable quantum processing on atom chips.     

利用mRNA差异显示技术筛选棉纤维发育相关基因

优良品种的选育对棉花生产有着其他技术不可替代的作用.研究细胞分化的内在机理,可为通过遗传工程途径人为控制棉纤维生长发育,选育优良农艺性状的新种质提供依据[1].     

Golgi duplication in Trypanosoma brucei requires Centrin2

Centrins are highly conserved components of the centrosome, which in the parasitic protozoan T. brucei comprises the basal body and nucleates the flagellum used for locomotion. Here, we found TbCentrin2 in an additional bi-lobed structure near to the Golgi apparatus. One lobe was associated with the old Golgi, and the other became associated with the newly forming Golgi as the cell grew. Depletion of TbCentrin1 inhibited duplication of the basal body, whereas depletion of TbCentrin2 also inhibited duplication of the Golgi. Thus, a Centrin2-containing structure distinct from the basal body appears to mark the site for new Golgi assembly.     

乙烯利调控甘蔗基因差异表达研究

【目的】建立并优化乙烯利调控甘蔗基因差异表达的cDNA—AFLP分析体系和银染体系,利用该技术对乙烯利处理的甘蔗进行基因差异表达研究,探讨乙烯利调控甘蔗生长的分子机理。【方法】以甘蔗品种ROC16为材料,在甘蔗生长40d左右时,叶面喷施200mg／L乙烯利溶液（对照为清水）0、3、6、12、24、48h后,分别取甘蔗叶片提取,总RNA,建立cDNA—AFLP分析体系和银染体系;利用193对引物组合对乙烯利处理和对照的甘蔗叶片RNA混合池的样品进行差异表达分析,对部分差异转录片段进行回收、克隆、反向Northern杂交验证、序列分析和功能分析等。【结果】经cDNA—AFLP体系反应扩增,乙烯利处理和对照每个样品的扩增条带数均在35～80之间,处理和对照之间条带多态性明显。在反向Northern杂交验证分析中,24个测试样品中有11个无差异,假阳性率高达46％;在乙烯利作用下,甘蔗的CHI、GST、ARP、LHC、核结合蛋白基因以及一批未知基因差异表达,其中CHI、GST,LHC和核结合蛋白基因是与促进植物的抗病抗逆、提高光合作用等密切相关的因子。【结论】cDNA—AFLP技术是研究乙烯利诱导甘蔗相关基因差异表达和调控的一种较有效方法,具有可行性。乙烯利可能是通过调控甘蔗体内GST、CHI、A脐口LHC等基因的表达,从而表现出促进甘蔗生长、提高光合作用、增强抗病抗逆性等生理效应。     

不同启动子调控的DREB1A基因对黄瓜的遗传转化

以黄瓜(CucumissativusL.)主栽品种长春密刺的子叶节为受体材料,采用农杆菌介导法,将分别由诱导型启动子rd29A和组成型启动子35S调控的转录因子DREB1A基因导入黄瓜,获得大量PCR阳性植株。统计结果表明:诱导型启动子rd29A调控的DREB1A基因抗性芽率和再生植株PCR阳性率明显高于组成型启动子35S。