Human Monocytes - CD14, CD16 - Ziegler-Heitbrock


NF beta A, a factor required for maximal interleukin-1 beta gene expression is identical to the ets family member PU.1. Evidence for structural alteration following LPS activation.


We have previously identified and characterized the macrophage-, neutrophil- and B cell-specific nuclear factor beta A (NF beta A), which is involved in transcriptional regulation of the interleukin-1 beta (IL-1 beta) gene. NF beta A binds to a highly conserved sequence element located 6 bp upstream of the TATA motif within the IL-1 beta promoter and is required for maximal expression of the IL-1 beta gene. Here we show that NF beta A is identical to the previously identified ets gene family member PU.1. The NF beta A binding element shares 100% sequence identity with a novel PU.1 binding element recently found in the immunoglobulin J-chain promoter. Methylation interference DNA footprinting data demonstrated that NF beta A and PU.1 make identical protein/DNA contacts. In vitro synthesized PU.1 possesses a mobility and binding specificity identical to NF beta A as determined by electrophoretic mobility shift analysis (EMSA). Antisera directed against amino acids 39-55 of PU.1 recognizes NF beta A in a manner indistinguishable from PU.1 in EMSA 'supershift' studies. NF beta A and PU.1 also possess similar protein structure as determined by proteolytic clipping bandshift analysis. Furthermore, we show that PU.1 is able to transactivate an NF beta A-dependent promoter when co-transfected into HeLa cells which lack PU.1/NF beta A. EMSA studies using recombinant TATA binding protein (TBP) and PU.1 suggest that PU.1 may induce assembly of a distinct TBP-dependent complex on the IL-1 beta promoter. Finally, immunohistochemical confocal laser scanning microscopy studies suggest that LPS stimulation of RAW macrophages induces a structural change in the N-terminal transcriptional activation domain of PU.1.

Authors: Buras, J.A., Reenstra, W.R., Fenton, M.J.
Journal: Mol. Immunol., 32: 541-554
Year: 1995
PubMed: Find in PubMed