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. 1991 Apr;65(4):1803–1811. doi: 10.1128/jvi.65.4.1803-1811.1991

Derivatives of Moloney murine sarcoma virus capable of being transcribed in embryonal carcinoma stem cells have gained a functional Sp1 binding site.

V E Prince 1, P W Rigby 1
PMCID: PMC239988  PMID: 1848307

Abstract

The long terminal repeat (LTR) sequences of Moloney murine leukemia virus and its closely related derivative Moloney murine sarcoma virus (Mo-MSV) are incapable of directing transcription in embryonal carcinoma (EC) stem cells. The myeloproliferative sarcoma virus, a derivative of Mo-MSV, has several point mutations in the LTR and is transcribed more efficiently to allow productive infection of F9 EC cells. One of these mutations, at -166 with respect to the transcriptional start, creates a consensus binding site for the well-characterized mammalian transcription factor Sp1. We used gel retardation assays to demonstrate that F9 EC cell extracts form several complexes with the myeloproliferative sarcoma virus sequence around -166. One of these complexes involves a murine Sp1-like protein, which has immunoreactivity, DNA binding specificity, and electrophoretic mobility equivalent to those of purified human Sp1 protein. An equivalent complex forms on the corresponding Mo-MSV sequence but with a fivefold-lower affinity. Consistent with these observations, introduction of the single point mutation at -166 into the Mo-MSV LTR, creating a consensus Sp1 binding site, increases expression in F9 EC cells sixfold.

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