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Volume 271, Number 26, Issue of June 28, 1996 pp. 15743-15752
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Octamer Binding Factors and Their Coactivator Can Activate the Murine PU.1 (spi-1) Promoter

(Received for publication, November 7, 1995, and in revised form, February 21, 1996)

Hematology/Oncology Division, Department of Medicine, Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts 02215

PU.1 (spi-1), a member of the Ets transcription factor family, is predominantly expressed in myeloid and B cells, activates many B cell and myeloid genes, and is critical for development of both of these lineages. Our previous studies (Chen, H. M., Ray-Gallet, D., Zhang, P., Hetherington, C. J., Gonzalez, D. A., Zhang, D.-E., Moreau-Gachelin, F., and Tenen, D. G. (1995) Oncogene 11, 1549-1560) demonstrate that the PU.1 promoter directs cell type-specific reporter gene expression in myeloid cell lines, and that PU.1 activates its own promoter in an autoregulatory loop. Here we show that the murine PU.1 promoter is also specifically and highly functional in B cell lines as well. Oct-1 and Oct-2 can bind specifically to a site at base pair 55 in vitro, and this site is specifically protected in B cells in vivo. We also demonstrate that two other sites contribute to promoter activity in B cells; an Sp1 binding site adjacent to the octamer site, and the PU.1 autoregulatory site. Finally, we show that the B cell coactivator OBF-1/Bob1/OCA-B is only expressed in B cells and not in myeloid cells, and that OBF-1/Bob1/OCA-B can transactivate the PU.1 promoter in HeLa and myeloid cells. This B cell restricted coactivator may be responsible for the B cell specific expression of PU.1 mediated by the octamer site.

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