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的作用。哺乳动物唾液酸化由于其参与重要的生物学过程，如病原体-宿主相互作用以及免疫和神经系统的功能，已成为深入研究的焦点。唾液酸化途径的缺陷与多种病理学有关，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学方法的局限性给阐明哺乳动物唾液酸化的生物学功能带来了重大困难。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。该系统的优点是基于其先进的遗传方法，丰富的信息，充分记录的发育事件，完整的基因组序列，和相对较低的遗传冗余。拟议的多学科研究的目的是在分子和遗传水平上对果蝇唾液酸转移酶基因进行全面表征，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物化学活性。为此，将在细胞培养物中表达唾液酸转移酶蛋白，使用亲和色谱法纯化，并测定其酶活性。另一个具体的目的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。为了精确地绘制唾液酸转移酶的表达，将在免疫染色和原位杂交分析中使用不同的分子标记。唾液酸转移酶的功能将在分子、细胞和生物体水平上通过对唾液酸转移酶基因相关表型的综合表征进行分析，这些表型是通过几种遗传技术获得的，包括基因靶向和RNAi方法。最后，在本项目的框架内，将使用基于蛋白质组学的方法鉴定唾液酸化的体内分子靶标。这项研究将阐明果蝇唾液酸化的分子机制和生物学作用，并将阐明唾液酸化在哺乳动物（包括人类）中的生物学功能。

项目成果

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

• 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.

Glycomic studies of Drosophila melanogaster embryos.

果蝇胚胎的糖组研究。

• DOI: 10.1007/s10719-006-6693-4
• 发表时间: 2006
• 期刊: Glycoconjugate journal
• 影响因子: 3
• 作者: North,SimonJ;Koles,Kate;Hembd,Caleb;Morris,HowardR;Dell,Anne;Panin,VladislavM;Haslam,StuartM
• 通讯作者: Haslam,StuartM

Sialyltransferase regulates nervous system function in Drosophila.

• DOI: 10.1523/jneurosci.5253-09.2010
• 发表时间: 2010-05-05
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Repnikova E;Koles K;Nakamura M;Pitts J;Li H;Ambavane A;Zoran MJ;Panin VM
• 通讯作者: Panin VM