In both animals and plants, the formation of a polar axis is one of the first steps during embryogenesis. Cell fate must then be specified and maintained along this axis in order for proper development to occur. In animals, inherited and somatic mutations in the genes that regulate this process can lead to many types of cancers. Therefore it is important to identify and understand the mechanisms by which these proteins act. We have identified a mutation in the Arabidopsis TOPLESS gene (tpl-1) that cause the transformation of the apical half of the embryo (the shoot) into a second basal half (the root). The apical half of early tpl- 1 embryos express shoot fate genes, but these are lost later in embryogenesis and replaced by the expression of root fate genes. The TPL protein shows structural similarity to known co-repressors, and tpl-1 can be suppressed by mutations in the putative co-activator HAG1, a GCN5-1ike protein. This has lead us to a hypothesis wherein TPL acts as a transcriptional co-repressor, whose role is to fix the identity of the shoot by repressing genes required for root formation in the apical half of the embryo. We propose to test this hypothesis and use TPL and HAG to identify other key regulators of embryonic polarity.
In AIM 1, we will test if TPL can act as a transcriptional repressor using both yeast and in planta transcriptional repression assays and determine if the tpl-1 mutation affects the ability of TPL to repress transcription. We will also fuse TPL to known activation and repression domains and analyze the phenotypic consequences in transgenic Arabidopsis.
In AIM2, we will use microarray analysis to compare wild-type, tpl-1 and tpl-1;hag1 mutant embryos to identify the common downstream genes of TPL and HAG1. We will use inducible forms of TPL, tpl-1 and HAG1 to place these downstream genes into a temporal context.
In AIM3 we will perform an enhancer, a suppressor, and a modular misexpression screen on tpl-1 to identify other genes in the TPL/HAG 1 pathway. These experiments will shed light on one of the mechanisms by which plants specify and maintain cell types during embryogenesis. It will allow us to compare the proteins and logic used in plants to those used in cognate processes in animal development, leading to a better understanding of both.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM072764-04
Application #
7275921
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Haynes, Susan R
Project Start
2004-09-27
Project End
2009-08-31
Budget Start
2007-09-01
Budget End
2008-08-31
Support Year
4
Fiscal Year
2007
Total Cost
$385,673
Indirect Cost
Name
Salk Institute for Biological Studies
Department
Type
DUNS #
078731668
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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Crawford, Brian C W; Sewell, Jared; Golembeski, Greg et al. (2015) Plant development. Genetic control of distal stem cell fate within root and embryonic meristems. Science 347:655-9
Krogan, Naden T; Hogan, Kendra; Long, Jeff A (2012) APETALA2 negatively regulates multiple floral organ identity genes in Arabidopsis by recruiting the co-repressor TOPLESS and the histone deacetylase HDA19. Development 139:4180-90
Smith, Zachery R; Long, Jeff A (2010) Control of Arabidopsis apical-basal embryo polarity by antagonistic transcription factors. Nature 464:423-6
Gallavotti, Andrea; Long, Jeff A; Stanfield, Sharon et al. (2010) The control of axillary meristem fate in the maize ramosa pathway. Development 137:2849-56
Pauwels, Laurens; Barbero, Gemma Fernandez; Geerinck, Jan et al. (2010) NINJA connects the co-repressor TOPLESS to jasmonate signalling. Nature 464:788-91
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Krogan, Naden T; Long, Jeff A (2009) Why so repressed? Turning off transcription during plant growth and development. Curr Opin Plant Biol 12:628-36
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Long, Jeff A; Ohno, Carolyn; Smith, Zachery R et al. (2006) TOPLESS regulates apical embryonic fate in Arabidopsis. Science 312:1520-3