Award details

Chromatin organisation and cell-type specific gene expression in the Arabidopsis root

Reference	BBS/E/J/0000A208
Principal Investigator / Supervisor	Professor Peter Shaw
Co-Investigators / Co-Supervisors
Institution	John Innes Centre
Department	John Innes Centre Department
Funding type	Research
Value (£)	24,895
Status	Completed
Type	Institute Project
Start date	26/08/2004
End date	25/08/2005
Duration	12 months

Abstract

Aim: We will investigate the biochemical basis of the link between chromatin organisation and cell fate. We have found, by 3D FISH, visible differences in the chromatin organisation at the GL2 locus between the two-epidermal cell types of the Arabidopsis root. These differences reflect the expression state of the GL2 locus in non-hair and hair cells, suggesting that GL2 expression is controlled by chromatin organisation. Strategies: Construction of transgenic lines. To analyse the chromatin composition of the two- epidermal cell types we will construct GFP transgenic lines that specifically express the GFP marker in hair cells and non-hair cells. The GFP marker will be nuclear localised by fusing it to the histone H2B. To generate the marker line expressing the GFP in non-hair cells we will use the GL2 promoter to drive the transcription of the histone H2B fused to GFP. For hair cells we will use the RHD6 promoter to drive the transcription of the histone H2B fused to GFP. Isolation of nuclei of the two-epidermal cell types. Based on the specificity of GFP expression of these transgenic lines we will be able to separate individually nuclei from the two epidermal cell types from the other types of cells that constitute the root by the use of a fluorescence activated cell sorter. DNase I protection assay. We will then prepare chromatin from non-hair and hair nuclei to map the DNase I hypersensitive sites present in the genomic region at and around the GL2 locus in the two cell types. Hypersensitive sites indicate those chromatin regions that are most accessible to non-histone DNA binding proteins and that are several orders of magnitude more susceptible to nuclease cleavage than chromatin that is not actively transcribed. This experiment will be important to give us a preliminary idea of which DNA sequences are the targets of transcriptional regulation and are potential site for chromatin remodelling.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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