Novel activator protein-2α splice-variants function as transactivators of the ovine placental lactogen gene

Sean W. Limesand, Russell V. Anthony

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

Activator protein-2 (AP-2) has been implicated as a transactivator of the human and ovine placental lactogen (oPL) genes. Transcriptional enhancement through an AP-2 cis-acting element has been described for other genes expressed in the placenta, but the AP-2 isoform enhancing expression is species dependent. Transactivation of the oPL minimal promoter (-124 bp to +16 bp) by AP-2 was confirmed by mutational analysis in transiently transfected human choriocarcinoma cells (BeWo). AP-2α was localized in ovine chorionic epithelial cells by immunohistochemistry and a 3-kb transcript was identified by Northern hybridization. Four nearly full-length AP-2 cDNAs were isolated from an ovine placenta cDNA library. Nucleotide sequencing these cDNAs revealed that the AP-2 mRNA expressed in the ovine placenta shares identity with human AP-2α, but variations in the predicted N-terminus were observed, and three unique AP-2α splice-variants were identified. Expression of AP-2α variants in HepG2 cells, devoid of endogenous AP-2, indicates that enhancement through the AP-2 element in the oPL gene minimal promoter was variant dependent. RNA transcripts for all of the ovine AP-2α splice-variants were confirmed in ovine placenta by RT-PCR, and homologs for two variants were found in human placenta. However, only one AP-2α transcript, which shares identity to Xenopus AP-2α, was expressed in BeWo cells. Immunoblot analysis confirmed AP-2α variants in ovine chorionic binucleate cell nuclear extracts, one of which migrates similar to the AP-2α variant identified in BeWo cell nuclear extracts. These data indicate the presence of new mammalian AP-2α splice-variants that augment transactivation of the oPL gene in ovine chorionic binucleate cells.

Original languageEnglish (US)
Pages (from-to)2390-2401
Number of pages12
JournalEuropean Journal of Biochemistry
Volume268
Issue number8
DOIs
StatePublished - May 20 2001
Externally publishedYes

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Keywords

  • Activator protein-2 (AP-2)
  • Ovine
  • Placental lactogen gene
  • Transcriptional regulation
  • Trophoblast

ASJC Scopus subject areas

  • Biochemistry

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