The goal of the proposed research is to identify pancreatic transcription factors that establish and maintain the developmental program specific for the exocrine tissue compartment of the pancreas. We will begin (Specific Aim 1) by identifying the DNA-binding transcription factors that mediate the acinar cell activity of the transcriptional enhancer of the gene for PTFl-p48, a critical transcriptional activator of acinar specific genes. To do this we will define the minimal functional enhancer and then map nuclear protein binding sites by DNase I footprinting in vitro and computer-based searches for evolutionarily conserved binding sequences characteristic of specific transcription factor families. We will confirm the relevance of the footprints and consensus binding sites by analyzing the effects of mutating the sites on the activity of the enhancer. We will identify candidate DNA-binding transcription factors of the exocrine pancreas by screening all members of seven major transcription factor families for expression in the developing and the mature exocrine pancreas by an RT-PCR protocol (Specific Aim 2). We will match individual pancreatic factors with potential p48 enhancer binding sites through the known consensus binding sequences of the family to which they belong. We will verify the match between factor and the enhancer site by testing whether the nucleotide sequence requirements for binding match the sequence requirements for activity in the enhancer and whether the factor can activate a reporter gene through the element in transfected cells. We will screen additional candidates by examining whether their expression occurs at an embryonic stage consistent with a developmental regulatory role. Finally, we will test the role of candidate factors by forced expression of dominant-negative and constitutively active chimeric forms of the factors in embryonic pancreatic rudiments capable of morphogenesis in culture (Specific Aim 3).

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK061220-03
Application #
6649810
Study Section
Special Emphasis Panel (ZDK1-GRB-4 (O1))
Program Officer
Serrano, Jose
Project Start
2001-09-30
Project End
2006-06-30
Budget Start
2003-09-01
Budget End
2004-06-30
Support Year
3
Fiscal Year
2003
Total Cost
$312,000
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Biochemistry
Type
Schools of Medicine
DUNS #
800771545
City
Dallas
State
TX
Country
United States
Zip Code
75390
Hess, David A; Strelau, Katherine M; Karki, Anju et al. (2016) MIST1 Links Secretion and Stress as Both Target and Regulator of the UPR. Mol Cell Biol :
Jiang, Mei; Azevedo-Pouly, Ana; Deering, Tye G et al. (2016) MIST1 and PTF1 Collaborate in Feed-forward Regulatory Loops that Maintain the Pancreatic Acinar Phenotype in Adult Mice. Mol Cell Biol :
Hoang, Chinh Q; Hale, Michael A; Azevedo-Pouly, Ana C et al. (2016) Transcriptional Maintenance of Pancreatic Acinar Identity, Differentiation, and Homeostasis by PTF1A. Mol Cell Biol 36:3033-3047
Houghton, Jayne A L; Swift, Galvin H; Shaw-Smith, Charles et al. (2016) Isolated Pancreatic Aplasia Due to a Hypomorphic PTF1A Mutation. Diabetes 65:2810-5
Krah, Nathan M; Murtaugh, L Charles (2016) Differentiation and Inflammation: 'Best Enemies' in Gastrointestinal Carcinogenesis. Trends Cancer 2:723-735
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Hale, Michael A; Swift, Galvin H; Hoang, Chinh Q et al. (2014) The nuclear hormone receptor family member NR5A2 controls aspects of multipotent progenitor cell formation and acinar differentiation during pancreatic organogenesis. Development 141:3123-33
Direnzo, Daniel; Hess, David A; Damsz, Barbara et al. (2012) Induced Mist1 expression promotes remodeling of mouse pancreatic acinar cells. Gastroenterology 143:469-80
Afelik, Solomon; Qu, Xiaoling; Hasrouni, Edy et al. (2012) Notch-mediated patterning and cell fate allocation of pancreatic progenitor cells. Development 139:1744-53
Xuan, Shouhong; Borok, Matthew J; Decker, Kimberly J et al. (2012) Pancreas-specific deletion of mouse Gata4 and Gata6 causes pancreatic agenesis. J Clin Invest 122:3516-28

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