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Blood, Vol. 92 No. 10 (November 15), 1998:
pp. 3841-3847
AF4 Encodes a Ubiquitous Protein That in Both Native and MLL-AF4
Fusion Types Localizes to Subnuclear Compartments
Quanzhi Li,
Joy L. Frestedt, and
John H. Kersey
From the University of Minnesota Cancer Center and Departments of
Pediatrics and Laboratory Medicine/Pathology, University of Minnesota,
Minneapolis, MN.
Acute leukemia with t(4;11)(q21,q23) translocation results from the
in-frame fusion of the MLL to the AF4/FEL gene. In previous studies,
we and others demonstrated that AF4 transcripts are present in a
variety of hematopoietic and nonhematopoietic human cells. To
further study the wild-type and leukemia fusion AF4, we used glutathione S-transferase (GST)-fusion proteins as immunogens to produce rabbit polyclonal antibodies that were specific for normal
and chimeric AF4 proteins. Using Western blotting analysis, we
demonstrated that the AF4 gene encodes proteins with apparent molecular
weight of 125 and 145 kD. A 45-kD protein coprecipitated with AF4
protein in immunoprecipitation. Also, the anticipated MLL-AF4-encoded
240-kD protein was detected in all cell lines with t(4;11)
translocations; fusion proteins were present in lesser quantity than
the wild-type AF4. The proteins recognized by the antibodies are of the
predicted sizes of the AF4 and MLL-AF4-encoded proteins based on
previous DNA sequencing analysis. The MLL-AF4 fusion protein had a
similar subcellular distribution as AF4. Both t(4;11) and non-t(4;11)
leukemic cells showed a similar pattern of punctate nuclear staining in
all cell lines tested using confocal immunofluorescence microscopy. AF4
antibodies should be useful for further elucidation of the function of
AF4 in normal cellular physiology, as well as the function of MLL-AF4
in leukemogenesis. The antibodies should also be helpful for the
diagnosis of the MLL-AF4 fusion proteins in t(4;11) leukemias.
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