MECHANISMS INVOLVED IN THE INDUCTION OF FETAL HEMOGLOBIN

Project: Research project

Project Details

Description

Prior to becoming a full time faculty member at the University of South
Alabama, I completed four years of post-doctoral training, performing
research in the field of hemoglobin switching. The goal for my research
was the identification of new agents with fetal hemoglobin inducibility
for the treatment of sickle cell patients. One such agent, butyric acid
induces fetal hemoglobin both in vivo and in vitro. Recent data suggest
butyrate may require specific DNA sequences, in the promoter region of
several genes, in order to induce gene expression. The role of nuclear
trans-acting factors in this process has not been defined. With regards
to the effects of this agent in the gamma globin promoter, my studies
in transgenic mice demonstrated the inability of butyrate to reactive
a totally silenced gamma gene. Utilizing detailed truncation analysis
of the gamma globin promoter in transgenic animals, new insights were
gained into the cis sequences involved in the induction of gamma gene
expression by butyric acid. Subsequent studies in vitro in mouse
erythroleukemia cells suggested the sequence between nucleotides-822 and
-893 is necessary for modulation of gamma gene activity, following
induction with butyrate. In addition, gel shift analysis indicates
sequence specific proteins bind in this region. My hypothesis is that
butyrate induces gamma gene expression through modifications of sequence
specific nuclear proteins which bind in the upstream promoter region.
This hypothesis will be tested by completing the following Specific
Aims: 1) To establish a transient assay system to test the ability of
butyric acid to induce gamma gene expression in the presence of the
beta-globin gene. 2) To study the ability of the identified butyrate
response element to confer butyrate inducibility to a heterologous
promoter. 3) Isolation of the cDNA(s) encoding the butyrate response
element DNA-binding protein(s) using an in vivo one hybrid system. 4)
Establish a prokaryotic expression system for protein synthesis and
purification. 5) Define the characteristics of the sequence specific
protein by structure-function analysis. The information gained by this
analysis will impart new insights into the mechanism by which butyrate
induces gamma gene expression. My institution has provided an
environment conducive to strong basic research activities and sufficient
opportunity for meaningful collaborative efforts.
StatusNot started

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