The adenovirus major late promoter TATA box and initiation site are both necessary for transcription in vitro (original) (raw)

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Chiron Corporation

Emeryville, CA 94608, USA

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+Present address: Damon Biotech, 119 FOurth Avenue, needham Heights, MA 02194, USA

Author Notes

Revision received:

13 September 1984

Accepted:

13 September 1984

Published:

11 October 1984

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Michael F. Concino, Richard F. Lee, James P. Merryweather, Roberto Weinmann, The adenovirus major late promoter TATA box and initiation site are both necessary for transcription in vitro, Nucleic Acids Research, Volume 12, Issue 19, 11 October 1984, Pages 7423–7433, https://doi.org/10.1093/nar/12.19.7423
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Abstract

Mutagenized DNA templates and HeLa whole cell extracts were used to study the effects of promoter-specific base changes on invitro transcription. DNA templates where the initiating adenine (+1) was changed to thymidine (AT+1) 1n the adenovirus 2 major late transcription unit were transcribed with 50% efficiency of the unaltered template. We have described a mutant at the TATA box, where the A at position −28 was changed to a C (AC-28). Transcription efficiency was reduced to less than 20% of control in the AC-28 mutant (Concino et al., 1983, J. Biol. Chem. 258: 8493–8496). Primer extension analysis revealed increased 5′ end heterogeneity for invitro transcripts derived from AC-28 and AT+1 DNA templates. Specific transcription was completely abolished from AT+1 DNA templates when a second change was introduced within the TATA sequence, creating a double mutant (AC-28 . AT+1). Neither the AC-28 nor the AT+1 change alone had such an effect, suggesting a coordinated interaction in transcription initiation involving both the TATA box and the Initiation site.

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Author notes

+Present address: Damon Biotech, 119 FOurth Avenue, needham Heights, MA 02194, USA

© 1984 IRL Press Ltd.

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