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Nucleic Acids Research Advance Access originally published online on July 10, 2007
Nucleic Acids Research 2007 35(14):4882-4894; doi:10.1093/nar/gkm519
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Nucleic Acids Research, 2007, Vol. 35, No. 14 4882-4894
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Real-time monitoring of functional interactions between upstream and core promoter sequences in living cells of sea urchin embryos

Akiko Kobayashi1, Youko Watanabe1, Koji Akasaka2 and Tetsuro Kokubo1,*

1Division of Molecular and Cellular Biology, International Graduate School of Arts and Sciences, Yokohama City University, Yokohama, Kanagawa, 230-0045, Japan and 2Misaki Marine Biological Station, Graduate School of Sciences, University of Tokyo, 1024 Koajiro, Misaki, Miura, Kanagawa, 238-0225, Japan

*To whom correspondence should be addressed. Tel: 045-508-7237; Fax: 045-508-7369; Email: kokubo{at}tsurumi.yokohama-cu.ac.jp

Received May 24, 2007. Revised June 19, 2007. Accepted June 20, 2007.

There are some functional compatibilities between upstream and core promoter sequences for transcriptional activation in yeast, Drosophila and mammalian cells. Here we examined whether similar compatibilities exist in sea urchin embryos, and if so, whether they are dynamically regulated during early development. Two reporter plasmids, each containing a test promoter conjugated to either CFP or YFP, were concurrently introduced into embryos, and their expression patterns were studied by fluorescence microscopy. The upstream sequence of the Hemicentrotus pulcherrimus (Hp) OtxE promoter drives the expression of its own core promoter and that of Strongylocentrotus purpuratus (Sp) Spec2a in different embryonic regions, especially at the late gastrula stage. Interestingly, when the four putative transcription factor binding sites of this upstream sequence were individually mutated, the resulting sequences directed different spatiotemporal expression from the same set of two core promoters, indicating that combinations of upstream factors may determine core promoter usage in sea urchin embryos. In addition, the insertion or deletion of consensus or nonconsensus TATA sequences changed the expression profile significantly, irrespective of whether the upstream sequence was intact or mutated. Thus, the TATA sequence may serve as a primary determinant for core promoter selection in these cells.

Present address: Akiko Kobayashi, Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, 52-1 Yada, Suruga-Ku, Shizuoka 422-8526, Japan


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[Abstract] [Full Text] [PDF]


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