Genetics and Biochemistry of Sialylation in Drosophila

果蝇唾液酸化的遗传学和生物化学

• 批准号: 7942241
• 负责人: VLADISLAV M PANIN
• 金额: $ 10.96万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7942241
• 关键词: Adult; Affinity Chromatography; Animal Model; Animals; Antibodies; Attention; Axon; Biochemical; Biochemistry; Biological; Biological Assay; Biological Models; Biological Process; Brain; Cell Communication; Cell Culture Techniques; Cells; Data; Defect; Development; Developmental Process; Drosophila genus; Embryo; Engineering; Event; Fucose; Functional disorder; Gel; Gene Expression; Gene Proteins; Gene Targeting; Genes; Genetic; Genetic Techniques; Goals; Homologous Gene; Human; Immune; In Situ Hybridization; In Vitro; Interdisciplinary Study; Investigation; Lectin; Ligands; Light; Link; Mammals; Maps; Mass Spectrum Analysis; Messenger RNA; Molecular; Molecular Genetics; Molecular Target; Neoplasm Metastasis; Nervous System Physiology; Nervous system structure; Neurologic; Neuromuscular Diseases; Pathology; Pathway interactions; Pattern; Phenotype; Play; Polysaccharides; Procedures; Proteins; Proteomics; RNA Interference; Research; Role; Sialyltransferases; Signal Transduction; Staging; Synaptic plasticity; System; Time; Tissues; Visual system structure; base; gene function; genome sequencing; glycosylation; immune function; in vivo; molecular marker; mutant; neurogenesis; notch protein; pathogen; research study; scale up; sialylation; tumor; two-dimensional

DESCRIPTION (provided by applicant): The long-term goal of our research is to understand the role that plays glycosylation in regulating cell interactions during animal development. Mammalian sialylation has become the focus of intensive investigation because of its involvement in important biological processes, such as pathogen-host interactions and the functioning of the immune and nervous systems. Defects in the sialylation pathway have been implicated in multiple pathologies, including tumor metastases, impaired synaptic plasticity, and neuromuscular disorders. At the same time, the complexity of sialylation, and the limit on genetic approaches impose significant difficulties on elucidating biological functions of sialylation in mammals. This project is directed towards a comprehensive understanding of molecular and genetic mechanisms of sialylation in the Drosophila model system. The advantages of this system are based on its advanced genetic approaches, abundance of information on well-documented developmental events, the complete genome sequence, and relatively low genetic redundancy. The proposed multidisciplinary research is aimed at a comprehensive characterization of the Drosophila sialyltransferase gene at the molecular and genetic levels, as well as elucidating the role of sialylation in Drosophila development. One of the specific aims of this project is to comprehensively characterize the sialyltransferase biochemical activity. To this end, the sialyltransferase protein will be expressed in cell culture, purified using affinity chromatography, and assayed for its enzymatic activity. Another specific aim is to investigate in detail the expression pattern of the sialyltransferase (both gene and protein) during different developmental stages. To precisely map the expression of the sialyltransferase, different molecular markers will be used in immunostaining and in situ hybridization analyses. The function of the sialyltransferase will be analyzed at molecular, cellular and organismal levels by comprehensive characterization of the sialyltransferase gene-associated phenotypes obtained by several genetic techniques, including gene targeting and RNAi approaches. Finally, in the framework of this project, the in vivo molecular targets of sialylation will be identified using a proteomics- based approach. This proposed research will elucidate the molecular mechanism and biological role of sialylation in Drosophila and should shed light on biological functions of sialylation in mammals, including humans.

描述（由申请人提供）：我们研究的长期目标是了解糖基化在动物发育过程中调节细胞相互作用中的作用。哺乳动物唾液酸化已成为深入研究的焦点，因为它参与重要的生物过程，例如病原体与宿主的相互作用以及免疫和神经系统的功能。唾液酸化途径的缺陷与多种病理有关，包括肿瘤转移、突触可塑性受损和神经肌肉疾病。同时，唾液酸化的复杂性和遗传方法的限制给阐明哺乳动物唾液酸化的生物学功能带来了很大的困难。该项目旨在全面了解果蝇模型系统中唾液酸化的分子和遗传机制。该系统的优势在于其先进的遗传方法、详细记录的发育事件的丰富信息、完整的基因组序列以及相对较低的遗传冗余。拟议的多学科研究旨在在分子和遗传水平上全面表征果蝇唾液酸转移酶基因，并阐明唾液酸化在果蝇发育中的作用。该项目的具体目标之一是全面表征唾液酸转移酶的生化活性。为此，唾液酸转移酶蛋白将在细胞培养物中表达，使用亲和层析纯化，并测定其酶活性。另一个具体目标是详细研究唾液酸转移酶（基因和蛋白质）在不同发育阶段的表达模式。为了精确绘制唾液酸转移酶的表达图谱，将在免疫染色和原位杂交分析中使用不同的分子标记。通过多种遗传技术（包括基因靶向和 RNAi 方法）获得的唾液酸转移酶基因相关表型的综合表征，将在分子、细胞和生物水平上分析唾液酸转移酶的功能。最后，在该项目的框架内，将使用基于蛋白质组学的方法来确定唾液酸化的体内分子靶标。这项拟议的研究将阐明果蝇唾液酸化的分子机制和生物学作用，并应阐明哺乳动物（包括人类）唾液酸化的生物学功能。

项目成果

Glycomic studies of Drosophila melanogaster embryos.

果蝇胚胎的糖组研究。

• 发表时间: 2006-07
• 影响因子: 3
• 作者: North, Simon J;Koles, Kate;Hembd, Caleb;Morris, Howard R;Dell, Anne;Panin, Vladislav M;Haslam, Stuart M
• 通讯作者: Haslam, Stuart M

Sialylation in protostomes: a perspective from Drosophila genetics and biochemistry.

原口动物中的唾液酸化：果蝇遗传学和生物化学的视角。

• 发表时间: 2009-04
• 影响因子: 3
• 作者: Koles, Kate;Repnikova, Elena;Pavlova, Galina;Korochkin, Leonid I;Panin, Vladislav M
• 通讯作者: Panin, Vladislav M

Protein O-mannosylation in animal development and physiology: from human disorders to Drosophila phenotypes.

动物发育和生理学中的蛋白质 O-甘露糖基化：从人类疾病到果蝇表型。

• DOI: 10.1016/j.semcdb.2010.03.010
• 发表时间: 2010-08
• 期刊: Seminars in cell & developmental biology
• 影响因子: 7.3
• 作者: Nakamura, Naosuke;Lyalin, Dmitry;Panin, Vladislav M.
• 通讯作者: Panin, Vladislav M.

Sialyltransferase regulates nervous system function in Drosophila.

唾液酸转移酶调节果蝇的神经系统功能。

• 发表时间: 2010-05-05
• 作者: Repnikova, Elena;Koles, Kate;Nakamura, Michiko;Pitts, Jared;Li, Haiwen;Ambavane, Apoorva;Zoran, Mark J;Panin, Vladislav M
• 通讯作者: Panin, Vladislav M

The role of Drosophila cytidine monophosphate-sialic acid synthetase in the nervous system.

果蝇胞苷单磷酸唾液酸合成酶在神经系统中的作用。

• 发表时间: 2013-07-24
• 作者: Islam, Rafique;Nakamura, Michiko;Scott, Hilary;Repnikova, Elena;Carnahan, Mindy;Pandey, Dheeraj;Caster, Courtney;Khan, Saba;Zimmermann, Tina;Zoran, Mark J;Panin, Vladislav M
• 通讯作者: Panin, Vladislav M