The research proposed in this grant application is designed to elucidate the basic regulatory events that control the differentiation of mouse keratinocytes. The overall goal of this project is to test the hypothesis that Ca++ induced terminal differentiation of keratinocytes is brought about by changes in gene expression. Balb/MK keratinocytes will be used as the experimental cell system. cDNA sequences that represent the Ca++ induced differentiation gene will be cloned, isolated, and characterized. Functional analysis will seek to prove that specific mRNA species and/or its protein product is directly involved in the initiation of differentiation. Characterization of the expression of the putative genes that bring about differentiation will include: the kinetics and biology of expression, complete sequence, and the function of gene products (proteins) that control differentiation. A major goal is to demonstrate whether expression of the keratinocyte phenotype can be separated from terminal growth inhibition. The effects of transforming oncogenes on the induction of differentiation will be determined. Since the MK cell line is committed to keratinocyte lineage, the DNA from keratinocytes will be transfected into embryonic epithelial cells that are not committed to a developmental lineage in order to isolate the gene responsible for commitment to keratinocyte differentiation. A final goal of this application will be to investigate the cellular regulation of the gene(s) that control differentiation induced growth inhibition. Genomic clones of genes that regulate differentiation will be used to identify cis acting sequences and trans acting factors. It is proposed that an experimental system can be designed to analyze the Ca++ modulated switch leading to growth inhibition and differentiation. A possible two stage model for the differentiation of keratinocyte cells will be examined. The understanding of this control system -- stimulation/inhibition of proliferation and differentiation -- will allow a better understanding of the processes leading to neoplastic growth.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Program Projects (P01)
Project #
1P01CA048799-01A1
Application #
3817284
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Type
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37203
Satterwhite, D J; White, R L; Aakre, M E et al. (2001) TGF-beta1 regulates the expression of multiple max-interacting transcription factors in Balb/MK cells: implications for understanding the mechanism of action of TGF-beta1. Pediatr Res 50:67-75
Engel, M E; McDonnell, M A; Law, B K et al. (1999) Interdependent SMAD and JNK signaling in transforming growth factor-beta-mediated transcription. J Biol Chem 274:37413-20
Norgaard, P; Law, B; Joseph, H et al. (1999) Treatment with farnesyl-protein transferase inhibitor induces regression of mammary tumors in transforming growth factor (TGF) alpha and TGF alpha/neu transgenic mice by inhibition of mitogenic activity and induction of apoptosis. Clin Cancer Res 5:35-42
Lutterbach, B; Hann, S R (1999) c-Myc transactivation domain-associated kinases: questionable role for map kinases in c-Myc phosphorylation. J Cell Biochem 72:483-91
Gorska, A E; Joseph, H; Derynck, R et al. (1998) Dominant-negative interference of the transforming growth factor beta type II receptor in mammary gland epithelium results in alveolar hyperplasia and differentiation in virgin mice. Cell Growth Differ 9:229-38
Alexandrow, M G; Moses, H L (1998) c-myc-enhanced S phase entry in keratinocytes is associated with positive and negative effects on cyclin-dependent kinases. J Cell Biochem 70:528-42
Zhao, G Q; Liaw, L; Hogan, B L (1998) Bone morphogenetic protein 8A plays a role in the maintenance of spermatogenesis and the integrity of the epididymis. Development 125:1103-12
Engel, M E; Datta, P K; Moses, H L (1998) RhoB is stabilized by transforming growth factor beta and antagonizes transcriptional activation. J Biol Chem 273:9921-6
Datta, P K; Chytil, A; Gorska, A E et al. (1998) Identification of STRAP, a novel WD domain protein in transforming growth factor-beta signaling. J Biol Chem 273:34671-4
Serra, R; Johnson, M; Filvaroff, E H et al. (1997) Expression of a truncated, kinase-defective TGF-beta type II receptor in mouse skeletal tissue promotes terminal chondrocyte differentiation and osteoarthritis. J Cell Biol 139:541-52

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