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We have identified an in vivo footprint over the PuF
site on the translocated c-myc allele in Burkitt's
lymphoma cells. The PuF site on the silent normal c-myc allele
was unoccupied. We demonstrated by electrophoretic mobility shift
assay, electrophoretic mobility shift assay with antibody, UV
cross-linking followed by SDS-gel electrophoresis, and Western analysis
that Nm23H2 in B cell nuclear extracts bound to the c-myc PuF
site. Transfection experiments with c-myc promoter constructs
in both DHL-9 and Raji cells revealed that the PuF site functioned as a
positive regulatory element in B cells with a drop in activity with
mutation of this site. Access to this site is blocked in the normal
silent c-myc allele; these data suggest that the Nm23H2
protein is involved in deregulation of the translocated c-myc allele in Burkitt's lymphoma cells.
Volume 270,
Number 22,
Issue of June 2, pp. 13392-13398, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
s Lymphoma
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