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. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM069952-01
Application #
6720264
Study Section
Pathobiochemistry Study Section (PBC)
Program Officer
Marino, Pamela
Project Start
2004-09-01
Project End
2009-08-31
Budget Start
2004-09-01
Budget End
2005-08-31
Support Year
1
Fiscal Year
2004
Total Cost
$260,889
Indirect Cost
Name
Texas A&M University
Department
Biochemistry
Type
Schools of Earth Sciences/Natur
DUNS #
078592789
City
College Station
State
TX
Country
United States
Zip Code
77845
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Islam, Rafique; Nakamura, Michiko; Scott, Hilary et al. (2013) The role of Drosophila cytidine monophosphate-sialic acid synthetase in the nervous system. J Neurosci 33:12306-15
Nakamura, Naosuke; Stalnaker, Stephanie H; Lyalin, Dmitry et al. (2010) Drosophila Dystroglycan is a target of O-mannosyltransferase activity of two protein O-mannosyltransferases, Rotated Abdomen and Twisted. Glycobiology 20:381-94
Repnikova, Elena; Koles, Kate; Nakamura, Michiko et al. (2010) Sialyltransferase regulates nervous system function in Drosophila. J Neurosci 30:6466-76
Nakamura, Naosuke; Lyalin, Dmitry; Panin, Vladislav M (2010) Protein O-mannosylation in animal development and physiology: from human disorders to Drosophila phenotypes. Semin Cell Dev Biol 21:622-30
Koles, Kate; Repnikova, Elena; Pavlova, Galina et al. (2009) Sialylation in protostomes: a perspective from Drosophila genetics and biochemistry. Glycoconj J 26:313-24
Koles, Kate; Lim, Jae-Min; Aoki, Kazuhiro et al. (2007) Identification of N-glycosylated proteins from the central nervous system of Drosophila melanogaster. Glycobiology 17:1388-403
Luo, Yi; Koles, Kate; Vorndam, Wendy et al. (2006) Protein O-fucosyltransferase 2 adds O-fucose to thrombospondin type 1 repeats. J Biol Chem 281:9393-9
North, Simon J; Koles, Kate; Hembd, Caleb et al. (2006) Glycomic studies of Drosophila melanogaster embryos. Glycoconj J 23:345-54