AML-1, PU.1, and Sp3 regulate expression of human bactericidal/permeability-increasing protein.
Bactericidal/permeability-increasing protein (BPI) is an antimicrobial protein in neutrophils, stored in azurophil granules. Expression of BPI is absent in neutrophils of newborns and patients with secondary granule deficiency (SGD), possibly contributing to dysfunction of neutrophils. We report two alternative transcription start sites at 52 and 22bp upstream of the translation start. A proximal 222bp promoter conferring expression in myeloid cells was identified, and critical cis-acting sites for myeloid expression were contained within the 159bp upstream of translation start. Within this region, direct binding and transactivation by AML-1, PU.1, and Sp3 were demonstrated, as judged by electrophoretic mobility shift analysis. Moreover, transient transfections of C/EBPalpha or C/EBPepsilon to HeLa cells resulted in increased promoter activity, indicating a direct or indirect role for C/EBP. In conclusion, we provide evidence for AML-1, PU.1, and Sp3 cooperatively and directly mediating BPI-expression during myeloid differentiation.
Lennartsson
Andreas
A
Department of Hematology, Lund University, Lund, Sweden.
Pieters
Katrien
K
Ullmark
Tove
T
Vidovic
Karina
K
Gullberg
Urban
U
D023181:Antimicrobial Cationic Peptides; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics* / Q000378:metabolism*; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D050676:Core Binding Factor Alpha 2 Subunit; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics*; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D022423:Myeloid Cells / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D051705:Sp3 Transcription Factor; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D051742:Transcription Factor CHOP; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation / Q000235:genetics; D020298:U937 Cells; D000099232:Proto-Oncogene Protein Spi-1
D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D022762:CCAAT-Enhancer-Binding Proteins; D050676:Core Binding Factor Alpha 2 Subunit; D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins; D051705:Sp3 Transcription Factor; D015534:Trans-Activators; D051742:Transcription Factor CHOP; D014157:Transcription Factors; D000099232:Proto-Oncogene Protein Spi-1; C497249:DDIT3 protein, human; C493728:RUNX1 protein, human; C497121:SP3 protein, human; C021623:bactericidal permeability increasing protein