The role of transcription factor PU.1 in the activity of the intronic enhancer of the eosinophil-derived neurotoxin (RNS2) gene

Eosinophil-derived neurotoxin (EDN) found in the granules of human eosinophils is a cationic ribonuclease toxin. Expression of the EDN gene (RNS2) in eosinophils is dependent on proximal promoter sequences in combination with an enhancer located in the first intron. We further define here the active...

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Bibliographic Details
Published in:Blood 1998-03, Vol.91 (6), p.2126
Main Authors: van Dijk, T B, Caldenhoven, E, Raaijmakers, J A, Lammers, J W, Koenderman, L, de Groot, R P
Format: Article
Language:English
Subjects:
Binding Sites
Cell Lineage
DNA, Neoplasm - genetics
DNA, Neoplasm - metabolism
Enhancer Elements, Genetic
Eosinophil-Derived Neurotoxin
Eosinophils - metabolism
Gene Expression Regulation, Leukemic
HL-60 Cells
Humans
Introns - genetics
Neoplasm Proteins - physiology
Neurotoxins - biosynthesis
Neurotoxins - genetics
Proto-Oncogene Proteins - physiology
Ribonucleases
RNA, Messenger - biosynthesis
RNA, Messenger - genetics
RNA, Neoplasm - biosynthesis
RNA, Neoplasm - genetics
Trans-Activators - physiology
Transcription, Genetic
Transfection
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Summary:Eosinophil-derived neurotoxin (EDN) found in the granules of human eosinophils is a cationic ribonuclease toxin. Expression of the EDN gene (RNS2) in eosinophils is dependent on proximal promoter sequences in combination with an enhancer located in the first intron. We further define here the active region of the intron using transfections in differentiated eosinophilic HL60 cells. We show that a region containing a tandem PU.I binding site is important for intronic enhancer activity. This region binds multiple forms of transcription factor PU.I as judged by gel-shift analysis and DNA affinity precipitation. Importantly, introducing point mutations in the PU.I site drastically reduces the intronic enhancer activity, showing the importance of PU.I for expression of EDN in cells of the eosinophilic lineage.
ISSN:0006-4971