Gene Expression in Trichomonas vaginalis

描述（由申请方提供）：阴道毛滴虫是全球最常见的非病毒性传播感染的病原体，也是美国人群中最常见的寄生虫。增加传播艾滋病毒的妇女慢性感染这种寄生虫和最近增加的耐药毛滴虫病的发病率，保证更好地了解这种未充分研究的病原体的生物学。本研究旨在为T.流浪汉我们以前的基因表达分析导致了一系列的观察，揭示了这种寄生虫和它的人类宿主的转录机制之间的明显差异，这可能是用于药物设计。一个启动子元件，intitator（Inr），它定义了转录的起始位点，并指导RNA聚合酶II（RNAPII）到这个网站和一个新的转录因子（IBP 39），与Inr和RNAPII相互作用已被发现和表征。我们还发现T. vaginocytosis基因，并开发了体内测定，以表征这种原始真核生物中的剪接。本研究的具体目的是：（1）研究IBP 39与RNA聚合酶II（RNAPII）C端结构域（CTD）的相互作用;包括RNAPII前起始复合物（PIC）和与它们相互作用以协调转录起始的启动子基序的迷走神经蛋白;（3）确定毛滴虫中调节RNA剪接的结构特性和基序。这些研究将揭示用于调节原生生物、藻类、酵母和后生动物中基因表达的基本机制的保守和不同特征，从而为基因调控的进化提供见解。此外，它们将提供有关基因表达的关键特性的关键信息，这些信息可能导致可能的治疗应用。

DESCRIPTION (provided by the applicant): Trichomonas vaginalis is the etiologic agent of the most common non-viral sexually transmitted infection worldwide and is the most prevalent parasite found in the US population. Increased transmission of HIV in women chronically infected with this parasite and a recent increase in the incidence of drug resistant Trichomoniasis warrant a better understanding of the biology of this understudied pathogen. The proposed study aims to provide fundamental knowledge of the mechanisms underlying the regulation of gene expression in T. vaginalis. Our previous analyses of gene expression has led to series of observations that reveal distinct differences between the transcriptional machinery of this parasite and its human host that might be exploited for drug design. A promoter element, the intitator (Inr), which defines the start site of transcription and directs RNA polymerase II (RNAPII) to this site and a novel transcription factor (IBP39) that interacts with both the Inr and RNAPII has been discovered and characterized. We have also discovered the presence of introns in T. vaginalis genes and developed in vivo assays to characterize splicing in this primitive eukaryote. The specific aims of the proposed studies to continue this research are: (1) To characterize the interaction between IBP39 and the C-terminal domain (CTD) of RNA polymerase II (RNAPII), (2) To identify T. vaginalis proteins comprising the RNAPII pre-initiation complex (PIC) and the promoter motifs they interact with to coordinate initiation of transcription and (3) To define the structural properties and motifs that regulate RNA splicing in Trichomonas. These studies will reveal both conserved and divergent features of the basic machinery utilized to regulate gene expression in protists, Archaea, yeast and metazoa, providing insight into the evolution of gene regulation. Additionally, they will provide critical information on pivotal properties of gene expression that might lead to possible therapeutic applications.