Evidence for a stem cell-specific repressor of Moloney murine leukemia virus expression in embryonal carcinoma cells.
A negative regulatory element (NRE) spanning the tRNA primer-binding site (PBS) of Moloney murine leukemia virus (M-MuLV) mediates repression of M-MuLV expression specifically in embryonal carcinoma (EC) cells. We precisely defined the element by base-pair mutagenesis to an 18-base-pair segment of the tRNA PBS and showed that the element also restricted expression when moved upstream of the long terminal repeat. A DNA-binding activity specific for the M-MuLV NRE was detected in vitro by using crude EC nuclear extracts in exonuclease III protection assays. Binding was strongly correlated with repression in EC cells. Mutations within the NRE that relieved repression disrupted binding activity. Also, nuclear extracts prepared from permissive, differentiated EC cell cultures showed reduced binding activity for the NRE. These results indicate the presence of a stem cell-specific repressor that extinguishes M-MuLV expression via the NRE at the tRNA PBS.
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Central Research and Development, E. I. du Pont de Nemours & Co., Inc., Wilmington, Delaware 19880-0328.
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Scott
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D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005090:Exodeoxyribonucleases; D005786:Gene Expression Regulation; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics*; D009154:Mutation; D015345:Oligonucleotide Probes; D012343:RNA, Transfer / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping; D013724:Teratoma; D014162:Transfection
D015345:Oligonucleotide Probes; D012343:RNA, Transfer; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III