The Transcription Factor, Nm23H2, Binds to and Activates the Translocated c-myc Allele in Burkitt's Lymphoma

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...

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Bibliographic Details
Published in:The Journal of biological chemistry 1995-06, Vol.270 (22), p.13392-13398
Main Authors: Ji, Lin, Arcinas, Magdalena, Boxer, Linda M.
Format: Article
Language:English
Subjects:
Alleles
B-Lymphocytes - metabolism
Base Sequence
Binding Sites
Burkitt Lymphoma - genetics
Cell Line
DNA Primers
Genes, myc
Humans
Molecular Sequence Data
Monomeric GTP-Binding Proteins
NM23 Nucleoside Diphosphate Kinases
Nucleoside-Diphosphate Kinase
Transcription Factors - metabolism
Transcriptional Activation
Translocation, Genetic
Tumor Cells, Cultured
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Summary: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.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.270.22.13392