REGULATION OF THE COLONY STIMULATING FACTOR-1 GENE

Project: Research project

Description

Colony stimulating factor-1 (CSF-1; M-CSF) is required for the growth and
differentiation of macrophage progenitors as well as the survival of
mature macrophages. In contrast to the known biological effects of CSF-1,
little is known about the regulation of CSF-1 gene expression. The
objective of this proposal is to identify regulatory mechanisms operating
at the molecular and cellular level which modulate expression of the
mouse colony stimulating factor-1 gene in a fibroblastic and in a
monocyte/macrophage cell line. The goal of the first specific aim is to
compare the regulation of CSF-1 expression in these two cell lines, under
basal and stimulated conditions. Northern analysis of steady-state CSF-1
mRNA levels and nuclear run-on assays will be used to determine whether
expression of the CSF-1 gene is under transcriptional or post-
transcriptional control. The goal of the next three specific aims is to
created a molecular map of the mouse CSF-1 gene promoter region. First, a
physical map will be constructed of the 5' flanking region of the mouse
CSF-1 gene. This will be accomplished by restriction enzyme mapping,
subcloning and sequencing of a genomic fragment which includes sequences
that flank the 5' end of the transcribed portion of the CSF-1 gene.
Second, the transcriptional efficiency of the cloned regulatory regions
will be analyzed in the fibroblastic and monocyte/macrophage cell lines.
This will be accomplished by creating plasmid vectors that contain
portions of the 5' flanking material adjacent to the chloramphenicol
acetyltransferase gene, and using these vectors in transient transfection
assays to evaluate the transcriptional efficiency of the regulatory
region in the cell lines, under basal and stimulated conditions. Third,
cis-acting regulatory elements in the 5' flanking region of the mouse
CSF-1 gene will be identified. This will be accomplished by using
subcloned fragments shown to contain regulatory and monocyte/macrophage
cells in DNase 1 protection, gel retardation and methylation interference
assays. These studies should help to identify some of the mechanisms and
DNA sequences which are involved in regulating the expression of the CSF-
1 gene.
StatusFinished
Effective start/end date8/1/897/31/98

Funding

  • National Institutes of Health
  • National Institutes of Health: $76,185.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $138,224.00
  • National Institutes of Health: $225,752.00
  • National Institutes of Health: $208,962.00

Fingerprint

macrophage colony-stimulating factor
Macrophage Colony-Stimulating Factor
macrophages
genes
cell lines
Gene Expression
monocytes
Trans-Activators
gene expression
Fibroblasts
plasmid vectors
deoxyribonucleases
regulatory sequences
restriction mapping
methylation
promoter regions
gels
genomics

ASJC

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