Cells must be able to quickly respond to changes in their environment. An important aspect of many such responses is the fast and reversible modification of proteins to regulate their function. The transient phosphorylation of proteins through the combined action of kinases and phosphatases may be the most prominent example of such regulation. A similar, more recently discovered modification of proteins is the stabile addition of adenosine 5'--‐monophosphate group (AMP) to proteins. This novel regulatory mechanism has been called AMPylation after its initial discovery in the context of the bacterial VopS protein. Since then, bacterial and eukaryotic Fic domains have been found that can AMPylate proteins. Drosophila offers a significant advantage for the analysis of the physiological role of AMPylation because fly genomes encode only a single Fic domain protein.
The specific aims described in this proposal combine biochemical and genetic approaches in Drosophila to define the role that this mechanism plays in visual neurotransmission. This proposal aims (i) to determine the cellular sites of AMPylation activity using immunofluorescence and electron microscopy approaches, (ii) to define developmental stages and cell types that require AMPylation, (iii) to analyze the physiological consequences of AMPylation in the context of the Drosophila visual system, and (iv) to employ biochemical purification in conjunction with genetic interaction assays to identify the molecular targets regulated by AMPylation. Completion of these experiments will yield a comprehensive understanding of the signaling pathways in the visual system that are modified by AMPylation.

Public Health Relevance

AMPylation is the stabile addition of an adenosine 5'--‐monophosphate group (AMP) to proteins. Recent findings have revealed that this novel regulatory mechanism plays an important role in diverse physiological process in eukaryotic cells. This proposal combines biochemical and genetic approaches in Drosophila to define the role that this novel mechanism plays in visual neurotransmission. Understanding the mechanism by which AMPylation modifies photoreceptor responses is an important first step so that we can start to appreciate the potential of this novel layer of regulation in modulating visual processing in health and disease.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY021922-03
Application #
8531258
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Greenwell, Thomas
Project Start
2011-09-01
Project End
2016-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
3
Fiscal Year
2013
Total Cost
$302,100
Indirect Cost
$112,100
Name
University of Texas Sw Medical Center Dallas
Department
Neurosciences
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
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Casey, Amanda K; Moehlman, Andrew T; Zhang, Junmei et al. (2017) Fic-mediated deAMPylation is not dependent on homodimerization and rescues toxic AMPylation in flies. J Biol Chem 292:21193-21204
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Rahman, Mokhlasur; Ham, Hyeilin; Liu, Xinran et al. (2012) Visual neurotransmission in Drosophila requires expression of Fic in glial capitate projections. Nat Neurosci 15:871-5