The long term goal of the research is to understand the tissue and developmental-specific regulation of genes expressed in erythroid cells. The Investigators have concentrated on the regulation of genes in the b globin locus as a model of erythroid-specific gene expression. The regulation of these genes depends on transcription factors which are limited to erythroid cells or cells of closely related lineages, and on an organized chromatin structure dictated by the formation of hypersensitive sites within a region known as the LCR or locus control region. The investigations have led to the identification of a new cis element involved in the expression of the human b globin gene. This element confers a strong enhancer activity and appears to function in conjunction with hypersensitive site 2. Thus, the factor binding to this site is a candidate to mediate this critical interaction between the LCR and the globin promoter. In addition, it is known that transcription factors regulate the expression of other transcription factors and these interrelationships suggest a complex hierarchy of interaction that not only regulates specific gene products in a particular tissue, but also regulate each other to function in early development. It is thus likely that the study of the interrelationship among transcription factors will be important for understanding both the expression of these genes in adult tissue, and the differentiation of specific cells during early development. They have therefore also focused on the regulation of the EKLF erythroid-specific transcription factor gene.
The specific aims of our studies are as follows: 1) Identification and characterization of a new cis element in the distal 5' flanking sequence of the human b globin gene. We will characterize this site through footprinting and oligonucleotide competion studies and the factor that binds the element will be isolated and cloned with the use of the yeast one-hybrid system or conventional procedures. 2) Regulation of the EKLF gene. The expression of transcription factor genes is highly regulated and previous studies involving both erythroid and non-erythroid transcription factors are revealing interesting interrelationships. To understand this hierarchy, we plan to define the cis elements and transacting factors responsible for the tissue-specific expression of the EKLF gene. Together, these studies will provide new insights into mechanisms of gene regulation as well as the biology of hematopoietic systems.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK039585-10
Application #
2608419
Study Section
Hematology Subcommittee 2 (HEM)
Program Officer
Badman, David G
Project Start
1988-04-01
Project End
1999-02-02
Budget Start
1997-12-01
Budget End
1998-11-30
Support Year
10
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Cincinnati
Department
Genetics
Type
Schools of Medicine
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Anderson, K P; Crable, S C; Lingrel, J B (1998) Multiple proteins binding to a GATA-E box-GATA motif regulate the erythroid Kruppel-like factor (EKLF) gene. J Biol Chem 273:14347-54
Anderson, K P; Kern, C B; Crable, S C et al. (1996) Analysis of developmental switching in transgenic mice with 5' and 3' deletions in the human beta globin gene. Transgenic Res 5:245-55
Anderson, K P; Kern, C B; Crable, S C et al. (1995) Isolation of a gene encoding a functional zinc finger protein homologous to erythroid Kruppel-like factor: identification of a new multigene family. Mol Cell Biol 15:5957-65
Lloyd, J A; Case, S S; Ponce, E et al. (1994) Positive transcriptional regulation of the human gamma-globin gene. Gamma PE is a novel nuclear factor with multiple binding sites near the gene. J Biol Chem 269:19385-93
Boissy, R E; Boissy, Y L; Krakowsky, J M et al. (1993) Ocular pathology in mice with a transgenic insertion at the microphthalmia locus. J Submicrosc Cytol Pathol 25:319-32
Krakowsky, J M; Boissy, R E; Neumann, J C et al. (1993) A DNA insertional mutation results in microphthalmia in transgenic mice. Transgenic Res 2:14-20
Anderson, K P; Lloyd, J A; Ponce, E et al. (1993) Regulated expression of the human beta globin gene in transgenic mice requires an upstream globin or nonglobin promoter. Mol Biol Cell 4:1077-85
Lloyd, J A; Krakowsky, J M; Crable, S C et al. (1992) Human gamma- to beta-globin gene switching using a mini construct in transgenic mice. Mol Cell Biol 12:1561-7
Ponce, E; Lloyd, J A; Pierani, A et al. (1991) Transcription factor OTF-1 interacts with two distinct DNA elements in the A gamma-globin gene promoter. Biochemistry 30:2961-7
Krakowsky, J M; Panke, E S; Lee, R F et al. (1989) Analysis of possible repressor elements in the 5'-flanking region of the human beta-globin gene. DNA 8:715-21

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