| NSF Org: |
IOS Division Of Integrative Organismal Systems |
| Recipient: |
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| Initial Amendment Date: | August 13, 2001 |
| Latest Amendment Date: | March 23, 2005 |
| Award Number: | 0114965 |
| Award Instrument: | Continuing Grant |
| Program Manager: |
Jacqueline Banks
IOS Division Of Integrative Organismal Systems BIO Directorate for Biological Sciences |
| Start Date: | September 1, 2001 |
| End Date: | August 31, 2006 (Estimated) |
| Total Intended Award Amount: | $2,100,000.00 |
| Total Awarded Amount to Date: | $2,118,000.00 |
| Funds Obligated to Date: |
FY 2002 = $500,000.00 FY 2003 = $512,000.00 FY 2004 = $500,000.00 FY 2005 = $6,000.00 |
| History of Investigator: |
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| Recipient Sponsored Research Office: |
104 AIRPORT DR STE 2200 CHAPEL HILL NC US 27599-5023 (919)966-3411 |
| Sponsor Congressional District: |
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| Primary Place of Performance: |
104 AIRPORT DR STE 2200 CHAPEL HILL NC US 27599-5023 |
| Primary Place of
Performance Congressional District: |
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| Unique Entity Identifier (UEI): |
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| Parent UEI: |
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| NSF Program(s): |
PLANT FUNGAL & MICROB DEV MECH, DEVELOPMENTAL BIOLOGY CLUSTER |
| Primary Program Source: |
app-0102 app-0103 app-0104 app-0105 |
| Program Reference Code(s): |
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| Program Element Code(s): |
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| Award Agency Code: | 4900 |
| Fund Agency Code: | 4900 |
| Assistance Listing Number(s): | 47.074 |
ABSTRACT
Two-component systems are the primary means by which bacteria sense and respond to environmental stimuli. These systems are comprised of a number of distinct elements, namely histidine kinases, response regulators and in the case of phosphorelays, histidine phosphotransfer proteins (HPts). Genes encoding similar proteins to each of these elements have been identified in Arabidopsis, and for the majority of the 35 such genes no function has yet been definitively ascribed. An integrated approach to elucidate the function of these proteins in Arabidopsis is proposed. A combination of gene knockouts and inducible overexpression will be used to assess the roles of these genes in plant growth and development. The mutant plant lines will be characterized in terms of their response to biotic and abiotic factors such as hormones, light, and osmotic stress, and for their pattern of gene expression. Where in the plants these genes are expressed will be determined using a combination of GUS fusions and in situ RNA analysis. The location of the cognate proteins within the cell will also be delineated. To facilitate this localization, a series of 10 monoclonal antibodies will be generated to marker proteins, each of which resides on a distinct membrane. Protein complexes from Arabidopsis will be purified and analyzed to determine the interactions among these elements and to identify novel interacting proteins. Together, these studies will illuminate the signaling pathways in which each of these Arabidopsis two-component signaling elements function and how they interact to control plant growth and development.
The data from these studies will be deposited on a publicly accessible web page that is currently under construction at UNC (http://www.bio.unc.edu/research/two-component/). A link to this web site will be established on the TAIR database, and we will coordinate with TAIR to deposit data as appropriate. The knockout seeds will be made publicly available through deposition in the ABRC Stock Center at Ohio State. The monoclonals raised against the membrane marker proteins will be available for the cost of shipping through UNH and the cell lines will also be deposited with the American Type Culture Collection (ATCC).
In keeping with the goals of the 2010 project, functional analysis of the two-component signaling elements will aid in our understanding of plants at the organismal, cellular, and evolutionary levels. The research will provide functional information on the role of Arabidopsis two-component signaling systems in plant growth and development. The research will serve to define the subcellular location of the two-component signal transduction pathways, the interactions between the two-component signaling elements, and the downstream targets of the pathways. The research should clarify how a signal transduction mechanism that arose in bacteria has been adapted to plant signal transduction.
These studies should uncover the functions of several gene families in Arabidopsis. The proteins encoded by these gene families are predicted to interact and thus our studies should aid in the development of a paradigm for signaling specificity among interacting members of large gene families. In addition, tools will be developed that will be generally applicable in defining the subcellular location of proteins in Arabidopsis.
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