A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene

Hanna S. Radomska, Anne B. Satterthwaite, Natalie Taranenko, Sailaja Narravula, Diane S. Krause, Daniel G. Tenen

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Proper regulation of the human CD34 gene requires a combinatorial action of multiple proximal and long-range, cis elements. This report shows that, like the murine CD34 5' untranslated region (UTR), the corresponding region of the human CD34 gene is necessary for optimal promoter activity. We localized the most critical element of this region to base pairs +48/+75. Through oligonucleotide competition and antibody supershift experiments in electrophoretic mobility shift assays, we found that this sequence contains a binding site (CCAAT box) for the transcription factor NFY (nuclear factor Y), a factor mediating cell type-specific and cell-cycle regulated expression of genes. Mutating this site led to a 5-fold decrease in CD34 promoter activity in transient transfection experiments. Interestingly, NFY binds adjacently to the earlier identified c-myb binding site. Here we show that both binding sites are important for CD34 promoter function: mutating either site alone decreased CD34 promoter-driven reporter gene activity 4-fold. We also show that the integrity of the c-myb binding site is necessary for stabilization of NFY binding to its site. Such cooperation between c-myb, which is expressed in early hematopoietic cells, and NFY, which is expressed in many cell types, might contribute to specific activation of CD34 in stem cells. The CCAAT box motif was also noted in the 5' UTR of the murine CD34 gene, however, NFY did not bind to this region. Thus, our results indicate that the functional similarities between the human and murine CD34 5' UTRs are achieved through different molecular mechanism(s).

Original languageEnglish (US)
Pages (from-to)3772-3780
Number of pages9
JournalBlood
Volume94
Issue number11
StatePublished - Dec 1 1999

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Stem cells
Stem Cells
Genes
5' Untranslated Regions
Binding Sites
Electrophoretic mobility
Electrophoretic Mobility Shift Assay
Reporter Genes
Oligonucleotides
Base Pairing
Transfection
Assays
Cell Cycle
Transcription Factors
Stabilization
Experiments
Chemical activation
Cells
nuclear factor Y
Gene Expression

ASJC Scopus subject areas

  • Hematology

Cite this

Radomska, H. S., Satterthwaite, A. B., Taranenko, N., Narravula, S., Krause, D. S., & Tenen, D. G. (1999). A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene. Blood, 94(11), 3772-3780.

A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene. / Radomska, Hanna S.; Satterthwaite, Anne B.; Taranenko, Natalie; Narravula, Sailaja; Krause, Diane S.; Tenen, Daniel G.

In: Blood, Vol. 94, No. 11, 01.12.1999, p. 3772-3780.

Research output: Contribution to journalArticle

Radomska, HS, Satterthwaite, AB, Taranenko, N, Narravula, S, Krause, DS & Tenen, DG 1999, 'A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene', Blood, vol. 94, no. 11, pp. 3772-3780.
Radomska HS, Satterthwaite AB, Taranenko N, Narravula S, Krause DS, Tenen DG. A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene. Blood. 1999 Dec 1;94(11):3772-3780.
Radomska, Hanna S. ; Satterthwaite, Anne B. ; Taranenko, Natalie ; Narravula, Sailaja ; Krause, Diane S. ; Tenen, Daniel G. / A nuclear factor Y (NFY) site positively regulates the human CD34 stem cell gene. In: Blood. 1999 ; Vol. 94, No. 11. pp. 3772-3780.
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