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-Chain Gene Promoter1
*
Laboratory of Immunobiology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and
Laboratory of Immune Cell Biology, National Cancer Institute, and
Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892
In mature T cells, limited synthesis of the TCR-
subunit is
primarily responsible for regulating surface expression of TCRs.
Transcription of is directed by a complex promoter that includes
two potential binding sites for the Ets family of transcription factors
at -52 (zEBS1) and -135 (zEBS2). Mutation of these two sites results
in a marked reduction of transcription from this promoter. Using
electrophoretic mobility shift analysis, Elf-1 was demonstrated to be
the Ets family member that binds to these sites. One site, zEBS1,
matches the optimal Elf-1 consensus sequence in eight of nine bases,
making it the best match of any known mammalian Elf-1 binding site. A
role for Elf-1 in TCR- trans-activation was confirmed by
ectopic expression of Elf-1 in COS-7 cells. This resulted in an
increase in TCR- promoter activity that mapped to zEBS1 and zEBS2.
Additional support for the involvement of Elf-1 in TCR-
trans-activation derives from the finding that a GAL4-Elf-1
fusion protein trans-activated TCR- promoter constructs
that had been modified to contain GAL4 DNA binding sites. These results
demonstrate that Elf-1 plays an essential role in the
trans-activation of a constitutively expressed T
cell-specific gene, and that trans-activation occurs in the
context of the native promoter in both lymphoid and nonlymphoid cells.
Taken together with the existing literature, these data also suggest
that the requirement for inducible factors in Elf-1-mediated
trans-activation may decrease as the affinity and number of
Elf-1 sites increase.
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