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Cloning of the complete Ly-6E.1 gene and identification of DNase I
hypersensitive sites corresponding to expression in hematopoietic cells
AM Sinclair and EA Dzierzak
Department of Gene Structure and Expression, National Institute for Medical
Research, London, UK.
The Sca-1 antibody recognizes antigens encoded by members of the Ly-6
multigene family. These antigens are expressed on fetal and adult
hematopoietic stem cells, progenitor cells, mature activated T cells, and
some nonhematopoietic cells and are most likely encoded by the Ly- 6E.1 and
Ly-6A.2 genes. Characterization and isolation of regulatory elements of
Ly-6E.1 and A.2 genes that govern tissue-specific and high levels of
expression in the cells of the hematopoietic system (particularly stem
cells) are of considerable interest. To characterize the control elements
of this gene, we have cloned a 30-kb fragment encoding a fully functional
Ly-6E.1 gene and 13 kb of 5' and 13 kb of 3' flanking sequence.
Transfection studies in murine erythroleukemia (MEL) cells show that a
14-kb BamHI fragment from this clone is sufficient to confer Ly-6E.1 gene
expression at levels equivalent to those of the endogenous gene. By mapping
regions of chromatin sensitive to DNase I digestion, we have located
hypersensitive sites in the 5' and 3' regions of the gene in FDCP-1 cells,
MEL cells, and various T- cell lines. The appearance of two 5'
hypersensitive sites in hematopoietic cells correlates with Ly-6E.1
expression after gamma- interferon induction. We show that the presence of
hypersensitive sites in the 5' and 3' regions corresponds to Sca-1
expression, and we also discuss the localization of putative regulatory
control elements.
Volume 82,
Issue 10,
pp. 3052-3062,
11/15/1993
Copyright © 1993 by The American Society of Hematology
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