Nature 294, 178 - 180 (12 November 1981); doi:10.1038/294178a0

Point mutation in the TATA box curtails expression of sea urchin H2A histone gene in vivo

R. Grosschedl^*, B. Wasylyk^†, P. Chambon^† & M. L. Birnstiel^*‡

^*Institut für Molekularbiologie II der Universität Zürich, Hönggerberg, 8093 Zürich, Switzerland
^†Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 84 de Biologie Moléculaire et de Génie Génétique de L'INSERM, Institut de Chimie Biologique, Strasbourg, France
^‡To whom correspondence should be addressed.

In most, but not all, eukaryotic genes transcribed by polymerase B (II)^1–5, the site of initiation of transcription is preceded by a TATA^A _TA^A _T sequence⁴, commonly referred to as the Goldberg–Hogness⁶ or TATA box, which seems to be required for the generation of faithful 5' termini of mRNAs^7–14. For all genes except one^9,12,15–17, deletion of the TATA box reduces the rate of mRNA synthesis^{7,10,11,13,18–23}. Point mutations in the TATA box of the chick conalbumin gene diminish transcription in vitro 20–40-fold^21,24. The question remained of whether this effect would persist in vivo where far upstream sequences dramatically modulate gene expression^7–11,25,26. Unfortunately the chicken conalbumin gene is poorly expressed when introduced into eukaryotic cells in culture (B.W., unpublished results) or into Xenopus oocytes (S. Swany, personal communication). We have therefore constructed a new gene unit with a chimaeric promoter region by introducing the conalbumin wild-type or mutated TATA box region into the sea urchin H2A histone gene, which is known to be efficiently expressed in Xenopus oocytes^27–29. We report here that with both the wild-type and mutated chimaeric promoters, initiation of transcription occurs at a novel initiation site about 26 nucleotides downstream from the TATA or TAGA boxes, but in the mutant case, the amount of specific transcripts is decreased fivefold.

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