Notch is the best characterized of several Drosophila 'neurogenic' genes that control a wide range of cell fate determinations that start in the early embryo. Notch serves as a cell membrane-bound receptor. It is cleaved following cell-surface association with ligand, to produce a soluble, intracellular protein fragment that functions in nuclei. Our studies will be extended as follows: (a) Properties of constitutively active Notch receptors allow us to map metalloprotease Kuzbanian function during Notch proteolysis and signaling. We will characterize the step in Notch cleavage regulated by Kuzbanian, the subcellular location of Notch/Kuzbanian interaction, and the timing of Kuzbanian function in relation to other factors influencing Notch activity, especially Presenilin. (b) Two novel genetic screens will be employed to search for factors regulating Notch cleavage and signaling. Certain Notch transgenes produce dominant male lethality, and a selection for mutations that rescue this phenotype will be the basis for one strategy. A second genetic screen will involve the simultaneous over-expression of Delta and each of about2,000 randomly chosen target genes in a Notch-dependent developing tissue, the eye. (c) A cultured cell assay has been developed that allows signaling by various activated Notch proteins to be measured quantitatively and qualitatively. Proteins implicated by genetic screening will be directly tested in cultured cells for their correlated effects on a Notch-dependent transcriptional response, and Notch proteolysis. (d) We will determine the function of 4 newly identified Drosophila relatives of kuzbanian. One approach will involve a recently devised method for stably repressing each gene's function by tissue-specific expression of inverted-repeat RNAs. (e) Factors regulating Notch transport to and from the cell surface, and controlling nuclear translocation of Notch will be identified and characterized using certain transport deficient Notch proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025103-26
Application #
6684077
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Tompkins, Laurie
Project Start
1978-04-01
Project End
2005-12-31
Budget Start
2004-01-01
Budget End
2005-12-31
Support Year
26
Fiscal Year
2004
Total Cost
$367,400
Indirect Cost
Name
Rockefeller University
Department
Genetics
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
Lieber, Toby; Kidd, Simon; Young, Michael W (2002) kuzbanian-mediated cleavage of Drosophila Notch. Genes Dev 16:209-21
Kidd, Simon; Lieber, Toby (2002) Furin cleavage is not a requirement for Drosophila Notch function. Mech Dev 115:41-51
Wesley, C S; Saez, L (2000) Notch responds differently to Delta and Wingless in cultured Drosophila cells. J Biol Chem 275:9099-101
Wesley, C S; Saez, L (2000) Analysis of notch lacking the carboxyl terminus identified in Drosophila embryos. J Cell Biol 149:683-96
Wesley, C S (1999) Notch and wingless regulate expression of cuticle patterning genes. Mol Cell Biol 19:5743-58
Brennan, K; Tateson, R; Lieber, T et al. (1999) The abruptex mutations of notch disrupt the establishment of proneural clusters in Drosophila. Dev Biol 216:230-42
Kidd, S; Lieber, T; Young, M W (1998) Ligand-induced cleavage and regulation of nuclear entry of Notch in Drosophila melanogaster embryos. Genes Dev 12:3728-40
Neel, V A; Young, M W (1994) Igloo, a GAP-43-related gene expressed in the developing nervous system of Drosophila. Development 120:2235-43
Lyman, D; Young, M W (1993) Further evidence for function of the Drosophila Notch protein as a transmembrane receptor. Proc Natl Acad Sci U S A 90:10395-9
Lieber, T; Kidd, S; Alcamo, E et al. (1993) Antineurogenic phenotypes induced by truncated Notch proteins indicate a role in signal transduction and may point to a novel function for Notch in nuclei. Genes Dev 7:1949-65

Showing the most recent 10 out of 12 publications