Characterization and use of a novel AT-hook protein in Leishmania

利什曼原虫新型 AT-hook 蛋白的表征和应用

• 批准号: 7896108
• 负责人: Ashok A Aiyar
• 金额: $ 21.3万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-15 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7896108
• 关键词: AT Rich Sequence; AT-Hook Motifs; Acquired Immunodeficiency Syndrome; Affect; American; Amphotericin B; Anti-Retroviral Agents; Archaea; Area; Bacteria; Binding; Bioinformatics; Biology; Cell Cycle Regulation; Cell physiology; Chromatin; Chromatin Structure; Clinical; Code; Communicable Diseases; Country; Coupled; Crithidia; DNA; DNA biosynthesis; Development; Disease; Drug resistance; Drug toxicity; Effectiveness; Eukaryota; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Goals; Growth; HIV; HMGA1a Protein; Health; Homologous Gene; Human; In Vitro; Incidence; Infection; Leishmania; Leishmania major; Leishmaniasis; Lesion; Life Cycle Stages; Mammalian Cell; Military Personnel; Mining; Mitotic; Molecular; Molecular Genetics; Mus; Names; Nature; Nuclear; Parasites; Parasitic Diseases; Patients; Pattern; Pentamidine; Phlebotominae; Plasmid Cloning Vector; Plasmids; Process; Productivity; Property; Protein Binding; Proteins; Protozoa; RNA; Reagent; Research; Role; Toxic effect; Transcription Initiation; Transcriptional Regulation; Trypanosoma; Viral Proteins; Virus; Visceral; Visceral Leishmaniasis; Work; base; burden of illness; cost; design; effective therapy; fungus; inhibitor/antagonist; insight; macrophage; microbial; novel; novel therapeutics; pathogen; peptidomimetics; public health relevance; resistant strain; skin lesion; therapeutic target; therapy development; tool

DESCRIPTION (provided by applicant): Leishmaniasis is a debilitating infectious disease, with clinical manifestations ranging from skin lesions to fatal visceral infections. This sandfly-borne parasitic disease results from infection of human macrophages with trypanosomatid protozoan parasites belonging to the genus Leishmania. Current treatments for leishmaniasis are inadequate due to their high cost, low efficacy, toxicity, and the increasing incidence of drug-resistant strains. Consequently, it is vitally important to identify new targets for therapies against Leishmania. Using bioinformatic strategies, we have identified a class of Leishmania genes previously not studied in trypanosomatid protozoans. These genes code for AT-hook proteins, a class of proteins that participate in multiple cellular processes, including DNA replication, and transcription. The importance of these processes is indicated by their conservation across all species. DNA replication and transcription have served as targets for the development of anti-microbials, including anti-retroviral therapies against HIV to reduce disease burden in AIDS patients. The essential nature of DNA replication and transcription for the survival/replication of all pathogens renders them attractive targets against diseases associated with Leishmania infections. Therefore, elucidating the role of AT-hook proteins in Leishmania promises new avenues through which effective novel therapeutics against this protozoan parasite may be developed. As their name suggests, AT-hook proteins bind AT-rich DNA. Although Leishmania genomes are highly GC-rich, they contain AT-rich sequences, some of which lie in "strand-switch regions" that are associated with initiation of RNA transcription. Toward understanding the role of AT-hook proteins in the biology of Leishmania, we have chosen to study the Leishmania major gene, LmjF06.0270, that is predicted to encode a 180 kD AT-hook protein. LmjF06.0270 was selected because it is highly conserved in all Leishmania species, but is absent in other eukaryote species, including closely-related trypanosomatids such as Trypanosoma and Crithidia. Therefore, LmjF06.0270 represents a novel target against diseases caused by all species of Leishmania. In this application, we have proposed studies to characterize the molecular functions of LmjF06.0270. In addition, based on known properties of AT-hook proteins conserved from Archaebacteria to metazoan eukaryotes, we propose to exploit LmjF06.0270 to develop a novel tool that will significantly augment genetic and molecular studies in all Leishmania species. PUBLIC HEALTH RELEVANCE: Leishmaniasis threatens 350 million people worldwide, with 500,000 new cases of fatal visceral leishmaniasis each year. The WHO estimates that this often debilitating and fatal disease is widespread in 88 countries worldwide, affecting Americans working in endemic areas, including military personnel. The goal of this research is to develop therapies to reduce these numbers, thus alleviating suffering and promote human health and productivity.

描述（由申请人提供）：利什曼病是一种令人衰弱的感染性疾病，临床表现范围从皮肤病变到致命的内脏感染。这种沙蝇传播的寄生虫病是由属于利什曼原虫属的锥虫原生动物寄生虫感染的人类巨噬细胞的感染。目前对利什曼病的治疗不足，因为它们的成本高，低功效，毒性以及抗药性菌株的发病率不断增加。因此，确定针对利什曼尼亚的疗法的新靶标非常重要。使用生物信息学策略，我们已经确定了以前未在锥虫原生动物中研究的利什曼原虫基因。这些基因代码用于AT-HOC蛋白，这是一类参与多个细胞过程的蛋白质，包括DNA复制和转录。这些过程的重要性通过它们在所有物种中的保护表明。 DNA复制和转录已成为抗菌剂的发展的靶标，包括针对HIV的抗逆转录病毒疗法，以减轻艾滋病患者的疾病负担。 DNA复制和转录的生存/复制的基本性质使它们对与利什曼原虫感染相关的疾病的有吸引力的靶标。因此，阐明At-Hook蛋白在Leishmania中的作用有望通过新的途径，通过这种途径，可以开发出有效的针对这种原生动物寄生虫的新型疗法。顾名思义，At-Hook蛋白结合了富含AT的DNA。尽管Leishmania基因组高度富含GC，但它们包含富含GC的序列，其中一些序列位于与RNA转录启动有关的“ Strand-Switch区域”中。为了理解At-Hook蛋白在Leishmania生物学中的作用，我们选择研究Leishmania主要基因LMJF06.0270，该基因预计将编码180 kd的at-hook蛋白质。之所以选择LMJF06.0270，是因为它在所有利什曼原虫物种中都是高度保守的，但是在其他真核生物中不存在，包括密切相关的锥形剂，例如锥虫和克里氏菌。因此，LMJF06.0270代表了针对所有利什曼尼亚物种引起的疾病的新型靶标。在此应用中，我们提出了研究以表征LMJF06.0270的分子功能。此外，基于从考古细菌到后生真核生物保守的AT钩蛋白的已知特性，我们建议利用LMJF06.0270来开发一种新型工具，该工具将显着增强所有利什曼尼亚物种中的遗传和分子研究。 公共卫生相关性：利什曼病威胁全球3.5亿人，每年有50万例新的内脏利什曼病病例。世卫组织估计，在全球88个国家，这种经常使人衰弱和致命疾病广泛，影响了包括军事人员在内的流行地区工作的美国人。这项研究的目的是开发减少这些数字的疗法，从而减轻苦难并促进人类健康和生产力。