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Lineage-restricted regulation of the murine SCL/TAL-1 promoter
EO Bockamp, F McLaughlin, AM Murrell, B Gottgens, L Robb, CG Begley and AR Green
University of Cambridge, Department of Haematology, MRC Centre, UK.
The SCL/TAL-1 gene encodes a basic helix-loop-helix transcription factor
that is expressed in multipotent hematopoietic progenitors before lineage
commitment. Its expression is maintained during differentiation along
erythroid, mast, and megakaryocytic lineages, but is repressed after
commitment to nonexpressing lineages. To begin to address the molecular
mechanisms underlying this complex pattern of expression, we have studied
the regulation of the murine SCL promoter in erythroid and T-cell lines.
Analysis of the methylation and chromatin structure of the SCL promoter
region showed that SCL mRNA expression correlated with DNase hypersensitive
sites and methylation status of the promoter. Transient reporter assays
showed that promoter 1a was active in erythroid cells but not in T cells.
Sequences between - 187 and +26 were sufficient for lineage-restricted
activity of promoter 1a. A joint promoter construct containing both
promoter 1a and promoter 1b also exhibited lineage-restricted activity.
Conserved GATA (-37), MAZ (+242), and ETS (+264) motifs were all shown to
contribute to SCL promoter activity in erythroid cells, but several other
motifs were not required for full promoter activity. The pattern of
complexes binding to the +242 MAZ and +264 ETS sites were the same in
erythroid and T cells. However, GATA-1 bound the -37 GATA site in erythroid
cells, whereas in T cells GATA-3 was only able to bind weakly, if at all.
Moreover, GATA-1 but not GATA-2 or GATA-3 was able to transactivate SCL
promoter 1a in a T-cell environment. These results suggest that inactivity
of SCL promoter 1a in T cells reflected the absence of GATA- 1 rather than
the presence of trans-dominant negative regulators.
Volume 86,
Issue 4,
pp. 1502-1514,
08/15/1995
Copyright © 1995 by The American Society of Hematology
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