The proposed studies are aimed at understanding how a single morphogen initiates the complex hierarchy of events leading to axis specification during animal development. The morphogen under investigation, Dorsal, is a transcription factor that is distributed in a dorsoventral concentration gradient in the Drosophila blastoderm embryo. This factor functions as both an activator and a repressor of transcription to establish multiple domains of gene activity along the dorsoventral axis. The diverse functions of Dorsal are mediated by a network of interactions between the Dorsal rel homology domain (RHD), the Dorsal Cterminal domain (CTD), and many additional regulatory factors. One Dorsal-interacting protein that is essential for Dorsal-mediated repression is the corepressor Groucho. The first specific aim of this proposal is to illuminate the mechanism of Dorsal-mediated activation by characterizing two sets of mutations discovered in a recent systematic mutagensis of Dorsal. These two classes of mutations define two distinct surfaces on the RHD. Affinity chromatography will be employed to identify novel coactivators that mediate the function of the surface defined by the class II mutations. Both genetic and biochemical approaches will be employed to test the hypothesis that the surface defined by the class I mutations modulates the activity of the CTD. The second specific aim is to determine the mechanism of repression by Groucho. Previous studies have led to a hypothesis for Groucho-mediated long-range repression that involves the recruitment of Groucho by Dorsal, the subsequent recruitment of a histone deacetylase by Groucho, and finally the spreading of Groucho along the deacetylated template. Genetic analysis, structural studies, and chromatin immunoprecipitation experiments will be combined to test this hypothesis rigorously. ? The pathway regulating dorsoventral patterning in Drosophila is homologous to pathways regulating the vertebrate immune response and vertebrate pattern formation. Thus, these studies may lead to a better understanding of human immunological and developmental disorders. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM044522-16
Application #
7029673
Study Section
Development - 1 Study Section (DEV)
Program Officer
Haynes, Susan R
Project Start
1990-08-07
Project End
2008-03-31
Budget Start
2006-04-01
Budget End
2007-03-31
Support Year
16
Fiscal Year
2006
Total Cost
$334,332
Indirect Cost
Name
University of California Los Angeles
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Chambers, Michael; Turki-Judeh, Wiam; Kim, Min Woo et al. (2017) Mechanisms of Groucho-mediated repression revealed by genome-wide analysis of Groucho binding and activity. BMC Genomics 18:215
Kwong, Pak N; Chambers, Michael; Vashisht, Ajay A et al. (2015) The Central Region of the Drosophila Co-repressor Groucho as a Regulatory Hub. J Biol Chem 290:30119-30
Kuo, Dennis; Nie, Minghua; De Hoff, Peter et al. (2011) A SUMO-Groucho Q domain fusion protein: characterization and in vivo Ulp1-mediated cleavage. Protein Expr Purif 76:65-71
Ajuria, Leiore; Nieva, Claudia; Winkler, Clint et al. (2011) Capicua DNA-binding sites are general response elements for RTK signaling in Drosophila. Development 138:915-24
Winkler, Clint J; Ponce, Alberto; Courey, Albert J (2010) Groucho-mediated repression may result from a histone deacetylase-dependent increase in nucleosome density. PLoS One 5:e10166
Ratnaparkhi, Girish S; Duong, Hao A; Courey, Albert J (2008) Dorsal interacting protein 3 potentiates activation by Drosophila Rel homology domain proteins. Dev Comp Immunol 32:1290-300
Duong, Hao A; Wang, Cheng Wei; Sun, Y Henry et al. (2008) Transformation of eye to antenna by misexpression of a single gene. Mech Dev 125:130-41
Duong, Hao A; Nagaraj, Raghavendra; Wang, Cheng W et al. (2008) Non-cell-autonomous inhibition of photoreceptor development by Dip3. Dev Biol 323:105-13
Qiao, Feng; Harada, Bryan; Song, Haiyun et al. (2006) Mae inhibits Pointed-P2 transcriptional activity by blocking its MAPK docking site. EMBO J 25:70-9
Song, Haiyun; Nie, Minghua; Qiao, Feng et al. (2005) Antagonistic regulation of Yan nuclear export by Mae and Crm1 may increase the stringency of the Ras response. Genes Dev 19:1767-72

Showing the most recent 10 out of 11 publications