A minimal serpin promoter with high activity in haematopoietic progenitors and activated T cells.
Authors
Hampson, LynneHampson, Ian N
Babichuk, Charolyn K
Cotter, Laura A
Bleackley, R Chris
Dexter, T Michael
Cross, Michael A
Affiliation
CRC Department of Hematopoietic Cell and Gene Therapeutics, Paterson Institute for Cancer Research, Wilmslow Road, Manchester M20 4BX, UK. lynne.hampson@man.ac.ukIssue Date
2001
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INTRODUCTION: The serine protease inhibitor Serpin 2A is highly expressed in ex vivo bipotent granulocyte/macrophage progenitor cells and in cultured myeloid stem cells. The gene undergoes rapid down-regulation as these cells are induced to differentiate, and constitutive expression in cultured myeloid stem cells retards maturation. Serpin 2A is also expressed in T cells as a consequence of activation. We now report analysis of the upstream regulatory elements that control Serpin 2A transcription. MATERIALS AND METHODS: Using primer extension and rapid amplification of cDNA ends the transcription start site of the Serpin 2A gene was mapped, and a 1.2 Kb genomic upstream fragment cloned and sequenced. Promoter activity and protein binding of deletion and site-directed mutant constructs were analysed by transient transfection and by electrophoretic mobility shift assays. RESULTS: A minimal promoter fragment was identified with high activity dependent on NF-kappa and Moloney murine leukaemia enhancer factor LVa binding sites in both myeloid stem cells and activated T cells. NF-kappa was shown to be the main DNA binding protein in T cells, whereas that in haematopoietic stem cells appears to be novel. CONCLUSION: Serpin 2A promoter activity in T cells is due predominantly to NF-kappa binding to its consensus site. Activity in haematopoietic stem cells appears to be mediated by a novel protein, which recognises the NF-kappa consensus only in the context of flanking sequences. This concise regulatory element may be of potential value in gene therapeutic applications.Citation
A minimal serpin promoter with high activity in haematopoietic progenitors and activated T cells. 2001, 2 (3):150-60 Hematol. J.Journal
The Hematology JournalDOI
10.1038/sj/thj/6200102PubMed ID
11920240Type
ArticleLanguage
enISSN
1466-4860ae974a485f413a2113503eed53cd6c53
10.1038/sj/thj/6200102
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