REGULATION OF EXPRESSION OF HUMAN GM-CSF

Project: Research project

Description

Colony-stimulating factors (CSFs) are required for the growth and
maturation of normal myeloid precursor cells in vitro, and a body of
indirect evidence suggests that they are primary regulators of
granulopoiesis in man. Human colony-stimulating factors have been
difficult to characterize because until recently, only small amounts of
partially purified protein were available. Our laboratory has recently
purified a 22,000 MW human GM-CSF (granulocyte-macrophage
colony-stimulating factor) that stimulates proliferation of hematopoietic
progenitor cells and primes neutrophils for enhanced responsiveness to
physiologic stimuli. Having obtained both cDNA and genomic clones encoding this human GM-CSF, it
will now be possible to define the molecular mechanisms regulating its gene
expression. GM-CSF mRNA has been detected in activated T lymphocytes and
HTLV-infected T-lymophoblast cell lines, but not in resting T cells. The
first steps will be to look for other sources of GM-CSF mRNA and to fully
characterize the induction of GM-CSF mRNA expression seen with T cell
activation. Next, the regions of the GM-CSF gene required for gene
expression in activated T cells will be defined by making recombinant
constructions containing fragments of the genomic GM-CSF clone linked to an
enzyme marker and measuring gene expression. The precise DNA sequences
necessary for regulation of GM-CSF gene expression will be identified by
making a series of deletion mutants and by performing
oligonucleotide-directed site-specific mutagenesis. I will also examine
the role of trans-acting transcriptional activators on GM-CSF expression in
both HTLV-infected T cells and activated T cells. These investigations should provide fundamental information regarding the
expression of human cellular genes in physiologic and pathologic states.
They should also provide valuable insights into the regulation of
hematopoiesis and the process of neoplastic transformation.
StatusFinished
Effective start/end date7/1/866/30/91

Funding

  • National Institutes of Health
  • National Institutes of Health: $77,328.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

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Colony-Stimulating Factors
T-Lymphocytes
Messenger RNA
Clone Cells
Neoplastic Processes
Gene Expression
Granulocyte Colony-Stimulating Factor
Myeloid Cells
Site-Directed Mutagenesis
Genes
Neutrophils
Complementary DNA
Cell Proliferation

ASJC

  • Medicine(all)