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A cell proliferation-dependent multiprotein complex NC-3A positively
regulates the CD34 promoter via a TCATTT-containing element
D Perrotti, T Bellon, R Trotta, R Martinez and B Calabretta
Department of Microbiology and Immunology, Thomas Jefferson University,
Philadelphia, PA, USA.
The CD34 cell surface antigen is a glycoprotein expressed by hematopoietic
stem and progenitor cells and also by certain nonhematopoietic cell-types.
Because CD34 expression is regulated both at the transcriptional and the
posttranscriptional level, we attempted to identify factors that, by
interacting with the 5' flanking region of the human CD34 gene, may
regulate its promoter activity in proliferating hematopoietic cells. By
electrophoretic mobility shift assay, UV cross-linking and DNase I
footprinting analyses, we identified a multiprotein complex, designated
NC-3A, that specifically interacts with the CD34 promoter region from
nucleotides -375 to -351. Sequence analysis of this region revealed the
presence of a distinct motif, TCATTT. Chloramphenicol acetyl-transferase
assays used to assess promoter activity in transiently transfected cells
showed that this TCATTT-containing element, which is conserved in both the
human and the murine CD34 genes, mediates positive regulatory activity in
hematopoietic and nonhematopoietic cells, and acts as an enhancer when
placed upstream of a heterologous promoter. Moreover, loss of CD34 promoter
activity was caused by mutation of the TCATTT motif. In addition, the
interaction of the nuclear multiprotein complex NC-3A with this enhancer
element is proliferation-dependent. These data indicate that, although not
cell-type specific, the formation of a multiprotein complex NC-3A
interacting with the region from nucleotides -375 to 351 plays an important
role in controlling CD34 promoter activity in proliferating hematopoietic
cells.
Volume 88,
Issue 9,
pp. 3336-3348,
11/01/1996
Copyright © 1996 by The American Society of Hematology
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