Transcription is a critical component in the expression of genes. Here we describe the design and analysis of a potent core promoter, termed super core promoter 1 (SCP1), which directs high amounts of transcription by RNA polymerase II in metazoans. SCP1 contains four core promoter motifs-the TATA box, initiator (Inr), motif ten element (MTE) and downstream promoter element (DPE)-in a single promoter, and is distinctly stronger than the cytomegalovirus (CMV) IE1 and adenovirus major late (AdML) core promoters both in vitro and in vivo. Each of the four core promoter motifs is needed for full SCP1 activity. SCP1 is bound efficiently by TFIID and exhibits a high propensity to form productive transcription complexes. SCP1 and related super core promoters (SCPs) with multiple core promoter motifs will be useful for the biophysical analysis of TFIID binding to DNA, the biochemical investigation of the transcription process and the enhancement of gene expression in cells.
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We are grateful to C.Y. Lim, J.-Y. Hsu and D. Urwin for advice throughout this study. We thank J.-Y. Hsu, T. Yusufzai, B. Rattner, D. Urwin, D. Fyodorov, J. Theisen, T. Bretz and C.Y. Lim for critical reading of the manuscript. We thank C. Inouye and R. Tjian for providing the purified human TFIID, C.Y. Lim for the purified D. melanogaster TFIID, J.-Y. Hsu for the pGL3-Basic vector with a modified polylinker, E.-T. Wong and G. Wahl for the pOG33-SV40 b-gal cotransfection plasmid, and W. Cavenee and F. Furnari for the use of their luminometer. This work was supported by a grant from the US National Institutes of Health (GM041249) to J.T.K.