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Blood ; 91(12): 4554-60, 1998 Jun 15.
Article in English | MEDLINE | ID: mdl-9616151

ABSTRACT

The recently-identified Wiskott-Aldrich syndrome protein gene (WASP) is responsible for the Wiskott-Aldrich X-linked immunodeficiency as well as for isolated X-linked thrombocytopenia (XLT). To characterize the regulatory sequences of the WASP gene, we have isolated, sequenced and functionally analyzed a 1.6-Kb DNA fragment upstream of the WASP coding sequence. Transfection experiments showed that this fragment is capable of directing efficient expression of the reporter chloramphenicol acetyltransferase (CAT) gene in all human hematopoietic cell lines tested. Progressive 5' deletions showed that the minimal sequence required for hematopoietic-specific expression consists of 137 bp upstream of the transcription start site. This contains potential binding sites for several hematopoietic transcription factors and, in particular, two Ets-1 consensus that proved able to specifically bind to proteins present in nuclear extracts of Jurkat cells. Overexpression of Ets-1 in HeLa resulted in transactivation of the CAT reporter gene under the control of WASP regulatory sequences. Disruption of the Ets-binding sequences by side-directed mutagenesis abolished CAT expression in Jurkat cells, indicating that transcription factors of the Ets family play a key role in the control of WASP transcription.


Subject(s)
Gene Expression Regulation , Hematopoietic Stem Cells/physiology , Proteins/genetics , Base Sequence , HeLa Cells , Humans , Jurkat Cells , Molecular Sequence Data , Protein Biosynthesis , Transfection , Wiskott-Aldrich Syndrome/genetics , Wiskott-Aldrich Syndrome Protein , X Chromosome
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