O-GlcNAc is the covalent modification of proteins at serine/threonine amino acids by the sugar moiety Nacetylglucosamine. The O-GlcNAc modification is not elongated to more complex oligosaccharide structures and is found only on nuclear and cytoplasmic proteins. The modification is highly dynamic and responds to signals of the extracellular environment such hormones, stress, and nutrients. Previously, I demonstrated that 0-GlcNAc is critical for the proper progression of the cell cycle in eukaryotic cells;gain of function of either 0-GlcNAc transferase (OGT), the enzyme which adds the modification, or 0-GlcNAcase (OGA), the enzyme which removes the modification, causes mitotic exit delays. Furthermore, OGT and OGA can be found in a signaling complex with mitotic kinases such as Aurora Kinase B. The goal of this proposal is to investigate the interactions of OGT with mitotic structures such as the spindle and with mitotic proteins. We hypothesize that OGT forms complexes with various proteins during M phase progression that then targets OGT the spindle and midbody. I plan to use multiple techniques such as using FRAP (Fluorescent Recovery after Photo-bleaching) on GFP-OGT to measure the kinetics of OGT at spindle/midbody, quantitative proteomics to identify OGT targeting proteins, and finally identifying the function of Aurora Kinase B in targeting OGT to mitotic substrates and structures. Our expected contribution is significant because we expect to find insight into the regulation of OGT by mitotic targeting proteins and how these interactions are uncoupled in diseases such as cancer.

Public Health Relevance

Many solid tumors show aspects of aneuploidy, a phenotype characterized by multinucleated cells. Gain of function of the O-GlcNAc transferase enzyme causes aneuploidy. The research proposed in this grant will provide significant biological information on how 0-GlcNAc transferase regulates mitosis and how alterations in OGT targeting can lead to the development of cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Exploratory Grants (P20)
Project #
5P20GM104936-07
Application #
8534220
Study Section
Special Emphasis Panel (ZRR1-RI-B)
Project Start
Project End
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
7
Fiscal Year
2013
Total Cost
$218,572
Indirect Cost
$73,822
Name
University of Kansas
Department
Type
DUNS #
016060860
City
Kansas City
State
KS
Country
United States
Zip Code
66160
Tran, Pamela V; Sharma, Madhulika; Li, Xiaogang et al. (2014) Developmental signaling: does it bridge the gap between cilia dysfunction and renal cystogenesis? Birth Defects Res C Embryo Today 102:159-73
Cha, Jeeyeon; Bartos, Amanda; Park, Craig et al. (2014) Appropriate crypt formation in the uterus for embryo homing and implantation requires Wnt5a-ROR signaling. Cell Rep 8:382-92
Pierce, A N; Ryals, J M; Wang, R et al. (2014) Vaginal hypersensitivity and hypothalamic-pituitary-adrenal axis dysfunction as a result of neonatal maternal separation in female mice. Neuroscience 263:216-30
Kumar, T Rajendra (2014) The quest for male germline stem cell markers: PAX7 gets ID'd. J Clin Invest 124:4219-22
Rumi, M A Karim; Dhakal, Pramod; Kubota, Kaiyu et al. (2014) Generation of Esr1-knockout rats using zinc finger nuclease-mediated genome editing. Endocrinology 155:1991-9
Tan, Ee Phie; Villar, Maria T; E, Lezi et al. (2014) Altering O-linked ?-N-acetylglucosamine cycling disrupts mitochondrial function. J Biol Chem 289:14719-30
Saadi, Irfan; Das, Pragnya; Zhao, Minglian et al. (2013) Msx1 and Tbx2 antagonistically regulate Bmp4 expression during the bud-to-cap stage transition in tooth development. Development 140:2697-702
Dasouki, Majed J; Rafi, Syed K; Olm-Shipman, Adam J et al. (2013) Exome sequencing reveals a thrombopoietin ligand mutation in a Micronesian family with autosomal recessive aplastic anemia. Blood 122:3440-9
Taniguchi, Cullen M; Finger, Elizabeth C; Krieg, Adam J et al. (2013) Cross-talk between hypoxia and insulin signaling through Phd3 regulates hepatic glucose and lipid metabolism and ameliorates diabetes. Nat Med 19:1325-30
Abrahamson, Dale R; St John, Patricia L; Stroganova, Larysa et al. (2013) Laminin and type IV collagen isoform substitutions occur in temporally and spatially distinct patterns in developing kidney glomerular basement membranes. J Histochem Cytochem 61:706-18

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