Single stranded DNA Binding Proteins in Trypanosoma cru*

克鲁锥虫中的单链 DNA 结合蛋白*

• 批准号: 6619435
• 负责人: Noreen Williams
• 金额: $ 4.03万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6619435
• 关键词: DNA binding protein; SDS polyacrylamide gel electrophoresis; South America; Trypanosoma cruzi; affinity chromatography; confocal scanning microscopy; cooperative study; fluorescence microscopy; gel mobility shift assay; gene expression; genetic recombination; genetic regulation; genetic transcription; immunofluorescence technique; messenger RNA; northern blottings; nucleic acid repetitive sequence; protein degradation; protein localization; protein purification; protein sequence; protein structure function; protozoal genetics; reporter genes; southern blotting; western blottings

DESCRIPTION (provided by applicant): American trypanosomiasis, or Chagas' disease is a major public health problem among poor rural populations in Latin America, where 16-18 million persons suffer from this disease, and 90 million are at risk. Sharp differences in regulation of gene expression between trypanosomatids and their mammalian hosts have been described, suggesting this process as an interesting target for drug design. In these parasites, the presence of polycistronic transcription means that post-transcriptional regulation of gene expression is the main step in determining the abundance of mRNAs. Using nuclear protein fractions from Trypanosoma cruzi epimastigotes, the group in Uruguay demonstrated the formation of at least three complexes that specifically recognize two dinucleotide repeated sequences (poly[dT-dG] and poly[dC-dA]) which are abundant in the vicinity of coding regions of T. cruzi genes. The specificity and preliminary estimation of equilibrium constants of these complexes are in good agreement with reported data for gene regulatory proteins. The goal of this project is to determine whether these dinucleotide repeats and the proteins that bind to them have a role in regulation of gene expression, transcription or in selective transcript processing, degradation or stabilization or recombination. The specific aims of this FIRCA project are: 1. to characterize the proteins interacting with the cis target (purification and microsequencing), to determine the fine subcellular localization of these proteins by immunofluorescence microscopy and to determine if they show stage specific expression by western analysis. 2. to characterize the genes encoding these proteins (cloning, sequencing and analysis by southern blot and chromosomal location as well as northern blot analysis of their expression). 3. to examine the dinucleotide sequence for effect on expression of reporter constructs. The location and the minimum length of the repeat sequence and, if it corresponds, the stage at which expression is altered [posttranscriptional, translational, etc.] will be examined. These experimental approaches will lead to an understanding of the role of these dinucleotide repeated sequences and the function and significance of the proteins that specifically recognize them providing insights into the regulation of gene expression in T. cruzi.

描述(申请人提供)：美国锥虫病，或查加斯病 在拉丁语中，疾病是农村贫困人口的主要公共卫生问题 美国有1600万至1800万人患有这种疾病，9000万人 都处于危险之中。在基因表达调控方面的显著差异 锥虫和它们的哺乳动物宿主已经被描述，这表明 过程是药物设计的一个有趣的目标。在这些寄生虫中， 多顺反子转录的存在意味着转录后 基因表达的调控是决定基因丰度的主要步骤 MRNAs。利用克氏锥虫上鞭毛体的核蛋白组分， 乌拉圭的研究小组证实了至少三个复合体的形成。 特异性识别两个二核苷酸重复序列(Poly[DT-DG] 和Poly[DC-da])，它们在T. 克鲁兹基因。均衡的特殊性和初步估计 这些复合体的常数与已报道的基因数据吻合较好。 调节蛋白。这个项目的目标是确定这些 二核苷酸重复序列和与之结合的蛋白质在 基因表达、转录或选择性转录本的调控 加工、降解或稳定或复合。的具体目标 这项FIRCA项目是：1.研究与蛋白质相互作用的 顺式靶标(纯化和微测序)，以确定罚款 免疫荧光显微镜和免疫荧光技术对这些蛋白的亚细胞定位 通过Western分析确定它们是否表现出阶段特异性表达。2.至 描述编码这些蛋白质的基因(克隆、测序和 Southern印迹、染色体定位和Northern印迹分析 分析它们的表达)。3.检查二核苷酸序列以确定 对报告结构表达的影响。位置和最低要求 重复序列的长度，如果它对应，则为 表达改变[转录后、翻译等]将会是 检查过了。这些实验方法将导致对 这些二核苷酸重复序列的作用及其功能和意义 专门识别它们的蛋白质提供了对 克鲁兹毛滴虫基因表达的调控。

项目成果

Trypanosoma cruzi: molecular characterization of an RNA binding protein differentially expressed in the parasite life cycle.

克氏锥虫：在寄生虫生命周期中差异表达的 RNA 结合蛋白的分子特征。

• DOI: 10.1016/j.exppara.2007.03.010
• 发表时间: 2007
• 期刊: Experimental parasitology
• 影响因子: 2.1
• 作者: Pérez-Díaz,Leticia;Duhagon,MaríaAna;Smircich,Pablo;Sotelo-Silveira,José;Robello,Carlos;Krieger,MarcoAurelio;Goldenberg,Samuel;Williams,Noreen;Dallagiovanna,Bruno;Garat,Beatriz
• 通讯作者: Garat,Beatriz

The Trypanosoma cruzi nucleic acid binding protein Tc38 presents changes in the intramitochondrial distribution during the cell cycle.

克氏锥虫核酸结合蛋白 Tc38 在细胞周期中呈现线粒体内分布的变化。

• DOI: 10.1186/1471-2180-9-34
• 发表时间: 2009
• 期刊: BMC microbiology
• 影响因子: 4.2
• 作者: Duhagon,MaríaA;Pastro,Lucía;Sotelo-Silveira,JoséR;Pérez-Díaz,Leticia;Maugeri,Dante;Nardelli,SheilaC;Schenkman,Sergio;Williams,Noreen;Dallagiovanna,Bruno;Garat,Beatriz
• 通讯作者: Garat,Beatriz