Characterisation of the GRAF gene promoter and its methylation in patients with acute myeloid leukaemia and myelodysplastic syndrome.
We report the isolation of the 5' flanking region of GRAF (GTPase regulator associated with the focal adhesion kinase), previously described as a putative tumour suppressor gene of acute myelogenous leukaemia and myelodysplastic syndrome, and demonstrate its promoter activity in reporter gene assays. Two putative protein-binding sites are identified of which one was sensitive to CpG methylation. The suppressed GRAF expression could be restored in leukaemia cell lines by treatment with a demethylating agent and an inhibitor of histone deacetylases. In contrast to normal tissues, which tested negative for GRAF promoter methylation, 11 of 29 (38%) bone marrow samples from patients with acute myeloid leukaemia or myelodysplastic syndrome were positive.
Bojesen
S E
SE
Department of Clinical Biochemistry, Copenhagen University Hospital, Herlev, Denmark.
Ammerpohl
O
O
Weinhäusl
A
A
Haas
O A
OA
Mettal
H
H
Bohle
R M
RM
Borkhardt
A
A
Fuchs
U
U
D000208:Acute Disease; D001483:Base Sequence; D002648:Child; D020690:GTPase-Activating Proteins / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D007951:Leukemia, Myeloid / Q000235:genetics*; D008745:Methylation; D008969:Molecular Sequence Data; D009190:Myelodysplastic Syndromes / Q000235:genetics*; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction
C115971:ARHGAP26 protein, human; D020690:GTPase-Activating Proteins