Abstract

The long term objectives of this proposal are to understand how the T- box gene family of transcription factors regulate gene expression in the kidney and how they contribute to kidney development and function. The focus of the current proposal is TBX2 and TBX3, two closely related members of the T-box family that are highly expressed in the kidney. TBX3 mutations result in ulnar-mammary syndrome, a human disorder associated with renal malformations. Based on their sequence conservation and expression pattern, it is hypothesized that TBX2 and TBX3 either serve a similar function or act to differentially regulate the same target genes. The following complementary in vitro and in vivo approaches are designed to test these hypotheses.
AIM 1. TO DEFINE THE ROLE OF tbx2 and TBX3 in RENAL GENE REGULATION WE WILL (i) IDENTIFY DOWNSTREAM TARGETS BY MEASURING ALTERED GENE EXPRESSION IN STABLE TRANSFECTANTS AND (ii) DETERMINE IF TBX2 AND TBX3 FUNCTION MAY BE MODULATED BY INTERACTIONS WITH OTHER T-BOX PROTEINS. (i) Transfectants in which TBX2 or TBX3 expression has been either induced or inactivated will be analyzed using cDNA arrays. (ii) Structural studies have shown that at least one T-box protein binds DNA as a dimer. The ability of other T-box proteins expressed in the kidney to heterodimerize with TBX2 or TBX3 will be tested.
AIM 2. TO DETERMINE THE ROLE OF TBX2 AND TBX3 IN NEPHROGENESIS, WE WILL CHARACTERIZE PROTEIN EXPRESSION IN MURINE KIDNEYS AND ASSESS THE AFFECT OF ALTERING TBX2 EXPRESSION. The pattern of TBX2 and TBX3 protein expression in mature and immature kidneys will be important in determining whether these genes function cooperatively or antagonistically and will be analyzed by immunocytochemistry. Transgenics expression human TBX2 will be analyzed for alterations in gene expression or kidney morphology using cDNA arrays and other molecular and biochemical markers. The proposed studies will extend our knowledge of the T-box family of developmental regulatory proteins and the role these genes play in the kidney, as well as provide further insight into the molecular mechanisms governing the process of nephrogenesis and the maintenance of normal kidney function.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
7R01DK048796-07
Application #
6500644
Study Section
Pathology A Study Section (PTHA)
Program Officer
Wilder, Elizabeth L
Project Start
1995-08-01
Project End
2003-03-31
Budget Start
2001-11-01
Budget End
2002-03-31
Support Year
7
Fiscal Year
2001
Total Cost
$188,801
Indirect Cost

  Institution

Name
State University of New York at Buffalo
Department
Biochemistry
Type
Schools of Medicine
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260

  Publications

Zweier, Christiane; Sticht, Heinrich; Aydin-Yaylagul, Inci et al. (2007) Human TBX1 missense mutations cause gain of function resulting in the same phenotype as 22q11.2 deletions. Am J Hum Genet 80:510-7
Wong, Yong Wee; Schulze, Christian; Streichert, Thomas et al. (2007) Gene expression analysis of nuclear factor I-A deficient mice indicates delayed brain maturation. Genome Biol 8:R72
Butz, Nataliya V; Gronostajski, Richard M; Campbell, Christine E (2006) T-box proteins differentially activate the expression of the endogenous interferon gamma gene versus transfected reporter genes in non-immune cells. Gene 377:130-9
Butz, Nataliya V; Campbell, Christine E; Gronostajski, Richard M (2004) Differential target gene activation by TBX2 and TBX2VP16: evidence for activation domain-dependent modulation of gene target specificity. Gene 342:67-76
Murtagh, Janice; Martin, Finian; Gronostajski, Richard M (2003) The Nuclear Factor I (NFI) gene family in mammary gland development and function. J Mammary Gland Biol Neoplasia 8:241-54
Carlson, H; Ota, S; Campbell, C E et al. (2001) A dominant repression domain in Tbx3 mediates transcriptional repression and cell immortalization: relevance to mutations in Tbx3 that cause ulnar-mammary syndrome. Hum Mol Genet 10:2403-13
Sinha, S; Abraham, S; Gronostajski, R M et al. (2000) Differential DNA binding and transcription modulation by three T-box proteins, T, TBX1 and TBX2. Gene 258:15-29
Seppala, R; Lehto, V P; Gahl, W A (1999) Mutations in the human UDP-N-acetylglucosamine 2-epimerase gene define the disease sialuria and the allosteric site of the enzyme. Am J Hum Genet 64:1563-9
He, M l; Wen, L; Campbell, C E et al. (1999) Transcription repression by Xenopus ET and its human ortholog TBX3, a gene involved in ulnar-mammary syndrome. Proc Natl Acad Sci U S A 96:10212-7
Chaudhry, A Z; Vitullo, A D; Gronostajski, R M (1998) Nuclear factor I (NFI) isoforms differentially activate simple versus complex NFI-responsive promoters. J Biol Chem 273:18538-46

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