Isolation and Chracterization of the Human Syncytin Gene Promoter 109회 lab meeting 2004.03.27 Isolation and Chracterization of the Human Syncytin Gene Promoter 석사 2년 박은실

Syncytin Trophoblast – syncytiotrophoblast - cytotrophoblast Mi et al, 2000 - induction of syncytialization in these cells can be prevented by syncytin antiserum. Frendo et al, 2003 - cell fusion and differentiation of normal human trophoblast cells in culture are markedly inhibited by specific syncytin antisense oligonucleotides.

HERV-W env & syncytin Intact HERV-W on chromosome 7q21-7q22 have 100% identity to SYNCYTIN gene HERV-W is highly expressed in human placenta, and weaker expression is noted in human testes. Expression is not observed in other human tissues. little is known about the mechanisms regulating the transcription of the syncytin gene.

Nucleotide sequences of 5’-flanking region of HERV-W0

Schematic diagram of the human BAC clone RG083m05 from 7q21 to 7q22. 2073bp 26725 28795 TATA +51~+70 Complemetary PCR Poly(a) RNA Mung Bean nuclease Protection Analysis

Transcription initiation site 30% decrease 50% decrease 90% decreasse Deletion analysis of the 59-flanking region of the HERV-W promoter in normal trophoblast cells.

DNAse 1 footprinting

DNase I footprint analysis of the proximal promoter region of the syncytin gene.

Site-Directed Mutagenesis

Gel shift and supershift assay

Gel shift and supershift analysis of FP2

Discussion Although full-length elements of HERVs are noninfectious, noninfectious, they have retained functional transcriptional elements, such as promoter and enhancer sequences, within their LTRs Sequence analysis of the 5’-flanking region of the HERV-W gene revealed a putative CCAAT box and many putative cis-acting elements the transcriptional regulation of the syncytin gene in trophoblast is highly dependent on the CCAAT motif and Oct site clearly demonstrate striking differences between the 3’-LTR and the 5’-LTR.