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 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.
van Dijk
T B
TB
Department of Pulmonary Diseases, University Hospital Utrecht, Utrecht, The Netherlands.
Caldenhoven
E
E
Raaijmakers
J A
JA
Lammers
J W
JW
Koenderman
L
L
de Groot
R P
RP
D001665:Binding Sites; D019070:Cell Lineage; D004273:DNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D047208:Eosinophil-Derived Neurotoxin; D004804:Eosinophils / Q000378:metabolism; D015973:Gene Expression Regulation, Leukemic; D018922:HL-60 Cells; D006801:Humans; D007438:Introns / Q000235:genetics*; D009363:Neoplasm Proteins / Q000502:physiology; D009498:Neurotoxins / Q000096:biosynthesis / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D012260:Ribonucleases; D015534:Trans-Activators / Q000502:physiology*; D014158:Transcription, Genetic; D014162:Transfection; D000099232:Proto-Oncogene Protein Spi-1
D004273:DNA, Neoplasm; D047208:Eosinophil-Derived Neurotoxin; D009363:Neoplasm Proteins; D009498:Neurotoxins; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D012260:Ribonucleases; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1; C479221:RNASE2 protein, human