Functional analysis of fish BCL-6 and Blimp-1 in vitro: Transcriptional repressors for B-cell terminal differentiation in fugu (Takifugu rubripes)

The transcriptional repressors BCL-6 and Blimp-1 are key regulators of B-cell terminal differentiation in mammals. We have previously identified the BCL-6 gene and Blimp-1 gene in fugu (Takifugu rubripes). In the present report, we conducted a functional analysis of fugu BCL-6 and Blimp-1 by using a one-hybrid reporter assay with Gal4 fusion proteins and Gal4DBD luciferase reporter gene. Results from the reporter assays in mammalian cell lines (HeLa, HEK-293, CV-1 and NIH3T3) and the fish cell line EPC show that Gal4-BCL6 and Gal4-Blimp1 strongly repress the transcription of the luciferase gene in all cell lines. Furthermore, deletion analyses show that the N-terminal region of BCL-6 has transcriptional repression activity; the BTB/POZ domain is an especially potent repression domain. In contrast to BCL-6, although the N-acidic domain and PR domain are insufficient for repression, most functional motifs of Blimp-1 are associated with transcriptional repression. These results suggest that BCL-6 and Blimp-1 are functional transcriptional repressors in fugu and that they regulate B-cell terminal differentiation in fugu.