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Sequences of the CyIIIa actin gene regulatory domain bound specifically by sea urchin embryo nuclear proteins

Thézé, Nadine and Calzone, Frank J. and Thiebaud, Pierre and Hill, Ronald L. and Britten, Roy J. and Davidson, Eric H. (1990) Sequences of the CyIIIa actin gene regulatory domain bound specifically by sea urchin embryo nuclear proteins. Molecular Reproduction and Development, 25 (2). pp. 110-122. ISSN 1040-452X. https://resolver.caltech.edu/CaltechAUTHORS:20151218-080936736

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Abstract

Expression of the Cylla cytoskeletal actin gene is a marker of differential gene activation in the aboral ectoderm of the early sea urchin embryo. Gene transfer expreiments have defined a 2,300 nucleotide cis-regulatory domain required for the correct spatial and temporal control of this gene. This domain includes at least 20 sites at which relatively stable DNA-protein complexes form in vitro on reaction with embryo nuclear extracts. We report the nucleotide sequence of the whole regulatory domain and map the sites at which high-specificity DNA-protein interactions occur. These were located initially by gel shift assays carried out on progressive restriction digests of given subfragments of the large regulatory domain and were located more exactly by oligonucleotide gel shift competitions. Eight of the sites of specific interaction are unique within the Cyllla regulatory domain, and the remainder consist of five different sites that occur more than once. We observe some well known sequences also found in regulatory regions of other genes, e.g., “CCAAT” and “octamer” elements. The various sites have been classified regarding putative biological function in other work, and the present studies permit an assessment of the number and complexity of interactions constituting each functional class and of the relative locations of sites of each class.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1002/mrd.1080250203DOIArticle
http://onlinelibrary.wiley.com/doi/10.1002/mrd.1080250203/abstractPublisherArticle
Additional Information:© 1990 Wiley-Liss, Inc. We thank Dr. Suzanna Horvath for oligonucleotide synthesis, and Dr. Christer Höög for assistance in ONP preparations. This work was supported by NIH grant HD-05753. P.T. was supported by a CNRS Fellowship,
Funders:
Funding AgencyGrant Number
NIHHD-05753
Centre National de la Recherche Scientifique (CNRS)UNSPECIFIED
Subject Keywords:DNA–protein interactions; Embryonic gene regulation; Regulatory factors
Issue or Number:2
Record Number:CaltechAUTHORS:20151218-080936736
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20151218-080936736
Official Citation:Thézé, N., Calzone, F. J., Thiebaud, P., Hill, R. L., Britten, R. J. and Davidson, E. H. (1990), Sequences of the CyIIIa actin gene regulatory domain bound specifically by sea urchin embryo nuclear proteins. Mol. Reprod. Dev., 25: 110–122. doi: 10.1002/mrd.1080250203
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:63056
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:26 Feb 2016 15:30
Last Modified:03 Oct 2019 09:24

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