Functional analysis of NTF-1, a developmentally regulated Drosophila transcription factor that binds neuronal cis elements

• Published in: Genes & development Vol. 3; no. 11; pp. 1677 - 1688
• Main Authors: DYNLACHT, B. D, ATTARDI, L. D, ADMON, A, FREEMAN, M, TJIAN, R
• Format: Journal Article
• Language: English
• Published: Cold Spring Harbor, NY Cold Spring Harbor Laboratory 01.11.1989
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; Cloning, Molecular; Diptera; DNA - metabolism; DNA-Binding Proteins - genetics; DNA-Binding Proteins - isolation & purification; DNA-Binding Proteins - metabolism; Drosophila - genetics; Drosophila Proteins; Drosophilidae; Fundamental and applied biological sciences. Psychology; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Neurons - metabolism; Nucleic Acid Hybridization; Promoter Regions, Genetic; Regulatory Sequences, Nucleic Acid; Transcription Factors - genetics; Transcription Factors - isolation & purification; Transcription Factors - metabolism; Transcription, Genetic; Transcription. Transcription factor. Splicing. Rna processing; Vaccinia virus - genetics; In vitro transcription; Embryonic development; Molecular hybridization; Purification; Insecta; Drosophila; In situ; Footprinting technique; DNA binding protein; Tissue specificity; Gene expression; Drosophilidae; Northern blotting; Arthropoda; DNA; Regulatory sequence; Invertebrata; Molecular cloning; Diptera; Aminoacid sequence
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.3.11.1677

In an effort to characterize sequence-specific transcription factors that regulate gene expression during Drosophila development, we identified and purified a novel DNA-binding activity (NTF-1). The purified protein consists of several polypeptides that bind selectively to a functionally important cis-control element of the Ultrabithorax (Ubx) promoter and to the neurogenic elements of both the dopa decarboxylase (Ddc) and fushi tarazu (ftz) promoter/enhancer regions. Purified NTF-1 activates transcription in vitro in a binding site-dependent manner through upstream sequences of the Ubx promoter. A cDNA clone encoding the open reading frame of NTF-1 was isolated, and the deduced primary amino acid sequence of NTF-1 includes a glutamine-rich region reminiscent of the transcriptional activation domains found in Sp1 but no recognizable DNA-binding domain. NTF-1 expression is temporally regulated during embryonic development. In addition, in situ hybridization experiments revealed that NTF-1 is transcribed in a spatially restricted pattern in the embryo, with the highest level of expression observed in the epidermis and a subset of cells in the CNS. Expression of the NTF-1 cDNA in mammalian cells yields a protein that displays DNA-binding and transcriptional activities indistinguishable from that of the collection of proteins isolated from Drosophila embryos. These findings suggest that NTF-1 is a member of a family of developmentally regulated transcription factors that may be involved in directing the expression of genes such as Ubx, Ddc, and ftz in neuronal cells.