Cell-restricted transcriptional modulators play critical roles in the process of selective gene regulation during hematopoiesis. We have been investigating the molecular and biological function of Erythroid Kr?ppel-like Factor (EKLF;KLF1). EKLF is a cell-restricted transcription factor that is essential for the erythroid program. Analysis of EKLF target gene regulation has revealed distinct transactivation mechanisms, and its protein interaction partners suggest additional unexplored roles in transcript control. The experiments of Aim 1 are directed at exploring the parameters of EKLF interactions with proteins and promoter DNA essential for these regulatory steps that lead to successful transcription. Analysis of EKLF's role in establishing an open chromatin structure, coupled with its interaction with histone H3 and the selective increase of H3.3 at the ?-globin promoter, suggest these observations are linked. As a result, the experiments of Aim 2 will address EKLF's role in coordinating histone H3.3 incorporation into chromatin. Analysis of the anemia in the heterozygous Nan mutant mouse has exposed an unexplained mechanism of gene-selective EKLF activation that leads to unique protein deficiency. The experiments of Aim 3 are designed to illuminate the molecular mechanism of the genetic distortion that results from the presence of Nan-EKLF in the erythroid cell. These studies will be aided by the use of in vivo assays, EKLF rescue systems, and primary or minimally manipulated cells. Elucidating EKLF's role in regulatory phenomena will continue to illuminate novel aspects of erythroid biology and the essential mechanisms by which a cell-restricted transcription factor can exert varied yet highly controlled influences on genetic expression and epigenetics.

Public Health Relevance

This proposal focuses on a continuing investigation of EKLF structure/function and how its protein-protein and protein-DNA interactions facilitate its ability o coordinate erythroid cell-specific control of chromatin modulation and gene transcription.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK046865-20
Application #
8389486
Study Section
Special Emphasis Panel (ZRG1-VH-D (02))
Program Officer
Bishop, Terry Rogers
Project Start
1993-08-01
Project End
2016-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
20
Fiscal Year
2012
Total Cost
$498,853
Indirect Cost
$197,410
Name
Icahn School of Medicine at Mount Sinai
Department
Biology
Type
Schools of Medicine
DUNS #
078861598
City
New York
State
NY
Country
United States
Zip Code
10029
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Gillinder, Kevin R; Ilsley, Melissa D; Nébor, Danitza et al. (2016) Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability. Nucleic Acids Res :
Yien, Yvette Y; Gnanapragasam, Merlin Nithya; Gupta, Ritama et al. (2015) Alternative splicing of EKLF/KLF1 in murine primary erythroid tissues. Exp Hematol 43:65-70
Siatecka, Miroslawa; Soni, Shefali; Planutis, Antanas et al. (2015) Transcriptional activity of erythroid Kruppel-like factor (EKLF/KLF1) modulated by PIAS3 (protein inhibitor of activated STAT3). J Biol Chem 290:9929-40
Lohmann, Felix; Dangeti, Mohan; Soni, Shefali et al. (2015) The DEK Oncoprotein Is a Critical Component of the EKLF/KLF1 Enhancer in Erythroid Cells. Mol Cell Biol 35:3726-38
Varricchio, Lilian; Dell'Aversana, Carmela; Nebbioso, Angela et al. (2014) Identification of NuRSERY, a new functional HDAC complex composed by HDAC5, GATA1, EKLF and pERK present in human erythroid cells. Int J Biochem Cell Biol 50:112-22
Manwani, Deepa; Bieker, James J (2014) KLF1: when less is more. Blood 124:672-3
Soni, Shefali; Pchelintsev, Nikolay; Adams, Peter D et al. (2014) Transcription factor EKLF (KLF1) recruitment of the histone chaperone HIRA is essential for β-globin gene expression. Proc Natl Acad Sci U S A 111:13337-42
Jaffray, Julie A; Mitchell, W Beau; Gnanapragasam, Merlin Nithya et al. (2013) Erythroid transcription factor EKLF/KLF1 mutation causing congenital dyserythropoietic anemia type IV in a patient of Taiwanese origin: review of all reported cases and development of a clinical diagnostic paradigm. Blood Cells Mol Dis 51:71-5
Yien, Yvette Y; Bieker, James J (2013) EKLF/KLF1, a tissue-restricted integrator of transcriptional control, chromatin remodeling, and lineage determination. Mol Cell Biol 33:4-13

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