The Drosophila eye is an excellent model system in which to study signaling pathways that are involved in the development of the human eye as well as in other developmental and disease contexts. A genetic mosaic screen has been used to identify novel genes required for the normal pattern of photoreceptor differentiation in the eye disc. This proposal will investigate several such genes that are required for normal activity of the Epidermal growth factor receptor (EGFR) signaling pathway, which controls the proliferation, survival and differentiation of retinal progenitor cells and is misregulated in many human cancers. One such gene, mago nashi, encodes a component of the exon junction complex (EJC), which is deposited onto all spliced mRNAs. Mutants lacking Mago nashi or other EJC subunits show a specific and dramatic reduction in the levels of the mRNA encoding Mitogen-activated protein kinase (MAPK), an essential downstream component of the EGFR and other receptor tyrosine kinase signaling pathways.
The first aim of this proposal is to determine whether the EJC is required for normal mapk pre-mRNA splicing, or regulates the stability of the mature mRNA. Either mechanism would represent a novel mode of action for this poorly understood complex, and would contribute to our understanding of the regulation of MAPK expression. Two additional genes required for photoreceptor differentiation encode endosomal proteins, Myopic and Vps4;
the second aim of this proposal will seek to understand the role of endocytosis in signaling by the EGFR and other receptors. In myopic mutant cells, EGFR protein accumulates but is unable to signal. The mechanism by which Myopic enhances EGFR signaling will be determined. The possible role in transcriptional regulation of a cleavage fragment of the EGFR formed within the endocytic pathway will also be investigated. The mutation in Vps4, which acts at the final step of protein sorting into multivesicular bodies for degradation, will be used as a tool to determine how receptor activity is altered during this sorting step.
The third aim will seek to clone and characterize two novel genes found in a screen of the X chromosome that are likely to affect signaling through the EGFR pathway. ayn and cassandra have been mapped to small chromosomal regions and will be identified by sequencing candidate genes. Biochemical methods will be used to determine their roles within each signaling pathway. Finally, mutations affecting CSN1b, a subunit of the COP9 signalosome, show a dramatic increase in the expression of the EGFR target gene argos. The hypothesis that this change is mediated by alterations in transcription factor ubiquitination will be investigated. Taken together, the proposal will reveal new molecular mechanisms in a conserved signaling pathway that is critical for retinal development.

Public Health Relevance

Retinal development is regulated by a conserved set of signaling pathways. Drosophila genetics offers a powerful, rapid and economical method to identify novel components required for this process and to understand their molecular mechanisms. We propose to characterize new genes required for signaling by the Epidermal growth factor receptor, which is essential for photoreceptor differentiation and the misregulation of which is a major cause of cancer. Evolutionary conservation suggests that our findings will be applicable to higher organisms, including humans, and may lead to tools for diagnosis or treatment of genetic disorders affecting the development of the eye or other organs, or to new cancer therapies.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY013777-12
Application #
8417702
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Neuhold, Lisa
Project Start
2002-02-01
Project End
2015-01-31
Budget Start
2013-02-01
Budget End
2015-01-31
Support Year
12
Fiscal Year
2013
Total Cost
$386,460
Indirect Cost
$158,460
Name
New York University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
Malone, Colin D; Mestdagh, Claire; Akhtar, Junaid et al. (2014) The exon junction complex controls transposable element activity by ensuring faithful splicing of the piwi transcript. Genes Dev 28:1786-99
Treisman, Jessica E (2013) Retinal differentiation in Drosophila. Wiley Interdiscip Rev Dev Biol 2:545-57
Steinhauer, Josefa; Liu, Hui Hua; Miller, Eli et al. (2013) Trafficking of the EGFR ligand Spitz regulates its signaling activity in polarized tissues. J Cell Sci 126:4469-78
Legent, Kevin; Steinhauer, Josefa; Richard, Magali et al. (2012) A screen for X-linked mutations affecting Drosophila photoreceptor differentiation identifies Casein kinase 1? as an essential negative regulator of wingless signaling. Genetics 190:601-16
Roignant, Jean-Yves; Legent, Kevin; Janody, Florence et al. (2010) The transcriptional co-factor Chip acts with LIM-homeodomain proteins to set the boundary of the eye field in Drosophila. Development 137:273-81
Roignant, Jean-Yves; Treisman, Jessica E (2010) Exon junction complex subunits are required to splice Drosophila MAP kinase, a large heterochromatic gene. Cell 143:238-50
Steinhauer, Josefa; Treisman, Jessica E (2009) Lipid-modified morphogens: functions of fats. Curr Opin Genet Dev 19:308-14
Roignant, Jean-Yves; Treisman, Jessica E (2009) Pattern formation in the Drosophila eye disc. Int J Dev Biol 53:795-804
Miura, Grant I; Roignant, Jean-Yves; Wassef, Michel et al. (2008) Myopic acts in the endocytic pathway to enhance signaling by the Drosophila EGF receptor. Development 135:1913-22
Legent, Kevin; Treisman, Jessica E (2008) Wingless signaling in Drosophila eye development. Methods Mol Biol 469:141-61

Showing the most recent 10 out of 18 publications