DEVELOPMENTALLY-REGULATED DNA-BINDING PROTEINS

Project Details

Description

The objective of this proposal is to elucidate the role of two DNA-binding
proteins in the development of a multicenter organism. ASF-1 and HSBF are
tobacco factors which bind to a hexameric motif present in the upstream
region of plant histone genes. This motif, TGACGT, is related to the
cAMP-responsive element in mammalian systems. ASF-1 and HSBF have different
sequence specificities as demonstrated by crosscompetition studies with
different binding site probes. In transgenic tobacco, recent data suggest
that the activity of a binding site specific for ASF-1 is induced by a
plant hormone and is also developmentally-regulated. Tobacco cDNA clones
that encode these two DNA-binding proteins have homologies to transcription
factors containing the leucine-zipper motif. Northern analyses show that
the expression of these genes is organspecific and
developmentally-regulated. Both proteins are encoded by small multigene
families. We want to characterize the regulation of the various members of
these two gene families. Since each individual member may be regulated
differently, we propose to isolate gene-specific probes from either the 5'
or 3' non-coding region of each member of these two gene families. These
probes will be used for nuclear run-on transcription assays to determine
whether the genes are regulated at the transcriptional level, and to
localize the mRNA of the particular gene to specific cell types by in situ
hybridization. The distribution of the gene products will also be examined
at the protein level by immune-detection techniques. The role of these gene
products in plant development will be studied by alterations of their
concentration in vivo through ectopic expression of the corresponding cDNA
in transgenic tobacco.
StatusFinished
Effective start/end date2/1/917/31/99

Funding

  • National Institutes of Health
  • National Institutes of Health: $95,400.00
  • National Institutes of Health
  • National Institutes of Health: $141,992.00
  • National Institutes of Health: $131,563.00
  • National Institutes of Health: $155,961.00
  • National Institutes of Health

ASJC

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)

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