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The Role of Transcription Factor PU.I in the Activity of the Intronic
Enhancer of the Eosinophil-Derived Neurotoxin (RNS2) Gene
Thamar B. van Dijk,
Eric Caldenhoven,
Jan A.M. Raaijmakers,
Jan-Willem J. Lammers,
Leo Koenderman, and
Rolf P. de Groot
From the Department of Pulmonary Diseases, University Hospital
Utrecht, Utrecht, The Netherlands.
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.
Blood, Vol. 91 No. 6 (March 15), 1998:
pp. 2126-2132
© 1998 by The American Society of Hematology.
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