An estrogen receptor beta isoform that lacks exon 5 has dominant negative activity on both ERalpha and ERbeta

An alternatively spliced isoform of human estrogen receptor beta (ERbeta) has been isolated from normal human testis mRNA that is coexpressed with wild-type ERbeta by reverse transcription polymerase chain reaction (RT-PCR). Sequence analysis of the ERbeta isoform PCR product reveals the absence of 139 bp that corresponds to the entire exon 5 of wild-type ERbeta, which predicts to lack part of the hormone-binding domain. The transient expression of the exon 5-deleted isoform of ERbeta (ERbetaDelta5) had no effect on basal transactivation activity of an estrogen-responsive luciferase reporter gene. This finding was in contrast to the previous reports that the exon 5-deleted isoform of ERalpha (ERalphaDelta5) acts as a dominant positive receptor, increasing basal gene transactivation itself. Moreover, when ERbetaDelta5 was cotransfected with the wild-type ERalpha or ERbeta, it behaved as a dominant negative receptor that inhibited not only estradiol-stimulated transactivation by ERbeta but also that by ERalpha. The ligand-independent nuclear localization of ERbetaDelta5 was confirmed by immunohistochemistry, and the coexpression of the isoform and the wild-type receptors could be observed in a single cell that transfected with both receptor cDNAs. These findings indicate that ERbetaDelta5 has a potential as a dominant negative receptor that blocks both ERalpha and ERbeta signaling pathways, suggesting some physiological roles of this isoform as an "ER inhibitor".