The overall goals of our program are to study in detail the properties and functional role of the lung Kruppel-like factor (LKLF/KLF2). This transcription factor is a member of a closely related family of KLF transcription factors. We have shown that LKLF in mice is necessary for embryos to survive past 12.5 days gestation and in addition have shown using chimeric animals that this transcription factor is necessary for normal lung development. We have also studied LKLF expression during development using whole mount tissue hybridization, and characterized its DNA binding and activating domains, as well as identified cis elements responsible for the regulated LKLF gene expression. The cis elements responsible for tissue specific expression of LKLF are conserved between mouse and human LKLF promoter. During the next granting period, we wish to further explore the inhibitory domain of LKLF. Interestingly, this domain inhibits transcription when bound to DNA next to a strong transcriptional activator. Preliminary studies suggest that a co-repressor exists and exerts its activity by binding to LKLF. This is demonstrated by the finding that overexpression of the inhibitory domain, without a DNA binding domain, alleviates the inhibition of LKLF presumably by sequestering a co-repressor. Using a two-hybrid yeast screen, we have identified a potential co-repressor molecule and this will be studied in detail during the next granting period. Modification of the inhibitory domain will be investigated including ubiquination and acetylation. In addition, we will investigate the regulation of the LKLF gene. We have already identified one region that is important for expression and will use transgenic mouse assays to define other elements. Once these elements are identified, the nuclear factors which bind to these elements will be defined, cloned and their activity studied. Finally, we will search for targets of the LKLF transcription factor. We will use gene microarray analysis of cells with and without the LKLF gene and develop conditional expression of LKLF to define downstream target genes in tissues such as lung, macrophages and T-lymphocytes.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL057281-08
Application #
6784568
Study Section
Lung Biology and Pathology Study Section (LBPA)
Program Officer
Berberich, Mary Anne
Project Start
1997-08-08
Project End
2006-07-31
Budget Start
2004-08-01
Budget End
2006-07-31
Support Year
8
Fiscal Year
2004
Total Cost
$341,007
Indirect Cost
Name
University of Cincinnati
Department
Genetics
Type
Schools of Medicine
DUNS #
041064767
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Pei, Liming; Leblanc, Mathias; Barish, Grant et al. (2011) Thyroid hormone receptor repression is linked to type I pneumocyte-associated respiratory distress syndrome. Nat Med 17:1466-72
Wu, Jinghai; Bohanan, Cynthia S; Neumann, Jon C et al. (2008) KLF2 transcription factor modulates blood vessel maturation through smooth muscle cell migration. J Biol Chem 283:3942-50
Huddleson, Justin P; Ahmad, Nisar; Lingrel, Jerry B (2006) Up-regulation of the KLF2 transcription factor by fluid shear stress requires nucleolin. J Biol Chem 281:15121-8
Ahmad, Nisar; Lingrel, Jerry B (2005) Kruppel-like factor 2 transcriptional regulation involves heterogeneous nuclear ribonucleoproteins and acetyltransferases. Biochemistry 44:6276-85
Wu, Jinghai; Srinivasan, Seetha V; Neumann, Jon C et al. (2005) The KLF2 transcription factor does not affect the formation of preadipocytes but inhibits their differentiation into adipocytes. Biochemistry 44:11098-105
Huddleson, Justin P; Ahmad, Nisar; Srinivasan, Seetha et al. (2005) Induction of KLF2 by fluid shear stress requires a novel promoter element activated by a phosphatidylinositol 3-kinase-dependent chromatin-remodeling pathway. J Biol Chem 280:23371-9
Wu, Jinghai; Lingrel, Jerry B (2005) Kruppel-like factor 2, a novel immediate-early transcriptional factor, regulates IL-2 expression in T lymphocyte activation. J Immunol 175:3060-6
Huddleson, Justin P; Srinivasan, Seetha; Ahmad, Nisar et al. (2004) Fluid shear stress induces endothelial KLF2 gene expression through a defined promoter region. Biol Chem 385:723-9
Wu, Jinghai; Lingrel, Jerry B (2004) KLF2 inhibits Jurkat T leukemia cell growth via upregulation of cyclin-dependent kinase inhibitor p21WAF1/CIP1. Oncogene 23:8088-96
Zhang, Xiaoling; Srinivasan, Seetha V; Lingrel, Jerry B (2004) WWP1-dependent ubiquitination and degradation of the lung Kruppel-like factor, KLF2. Biochem Biophys Res Commun 316:139-48

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