NF-Y binding is required for transactivation of neuronal aromatic L-amino acid decarboxylase gene promoter by the POU-domain protein Brn-2

Brain Research. Molecular Brain Research
C Dugast, M J Weber

Abstract

We have previously characterized binding sites for the NF-Y transcription factor (-71/-52) and Brn-2 POU-domain protein (-92/-71) in the neuronal promoter of the human aromatic L-amino acid decarboxylase gene [Mol. Brain Res. 56 (1998) 227]. We have now explored the functional role of these binding sites in transfected SK-N-BE neuroblastoma cells. Mutations of the NF-Y site that abolish binding depressed expression of a luciferase reporter gene up to 25-fold. The overexpression of a dominant negative mutant of NF-YA subunit depressed expression by 60%. Promoter activity was increased by the overexpression of Brn-2. Mutations or deletion of the binding site of Brn-2 did not suppress transcriptional activation by overexpressed Brn-2, while promoters defective in NF-Y binding were not transactivated by Brn-2. A GST-pulldown experiment showed that recombinant human Brn-2 protein weakly interacts with recombinant NF-Y outside of DNA. Cooperative binding of recombinant NF-Y and GST--Brn-2 proteins on the neuronal promoter was evidenced by an electrophoretic mobility shift assay. The POU-domain of Brn-2 was sufficient for such interaction. The results thus suggest that the activation of the neuronal promoter of the aromatic L-amino ac...Continue Reading

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Citations

May 2, 2006·Stem Cells and Development·Netta R BlondheimDaniel Offen
Nov 6, 2008·Pigment Cell & Melanoma Research·Anthony L Cook, Richard A Sturm
May 27, 2008·Molecular and Cellular Neurosciences·Lynn F SniderhanSanjay B Maggirwar
Dec 14, 2005·Journal of Molecular Biology·Sylvie BannwarthAnne Gatignol
Jun 22, 2002·Differentiation; Research in Biological Diversity·Xiaotao LiWilliam H Klein
Jan 8, 2019·American Journal of Medical Genetics. Part C, Seminars in Medical Genetics·Ni-Chung LeeWuh-Liang Hwu

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