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Published July 15, 2007 | public
Journal Article

Exclusive developmental functions of gatae cis-regulatory modules in the Strongylocentrorus purpuratus embryo

Abstract

The gatae gene of Strongylocentrotus purpuratus is orthologous to vertebrate gata-4,5,6 genes. This gene is expressed in the endomesoderm in the blastula and later the gut of the embryo, and is required for normal development. A gatae BAC containing a GFP reporter knocked into exon one of the gene was able to reproduce all aspects of endogenous gatae expression in the embryo. To identify putative gatae cis-regulatory modules we carried out an interspecific sequence conservation analysis with respect to a Lytechinus variegatus gatae BAC, which revealed 25 conserved non-coding sequence patches. These were individually tested in gene transfer experiments, and two modules capable of driving localized reporter expression in the embryo were identified. Module 10 produces early expression in mesoderm and endoderm cells up to the early gastrula stage, while module 24 generates late endodermal expression at gastrula and pluteus stages. Module 10 was then deleted from the gatae BAC by reciprocal recombination, resulting in total loss of reporter expression in the time frame in which it is normally active. Similar deletion of module 24 led to ubiquitous GFP expression in the gastrula and pluteus. These results show that Module 10 is uniquely necessary and sufficient to account for the early phase of gatae expression during endomesoderm specification. In addition, they imply a functional cis-regulatory module exclusion, whereby only a single module can associate with the basal promoter and drive gene expression at any given time.

Additional Information

© 2007 Elsevier Inc. Received for publication 10 February 2007; revised 1 May 2007; accepted 4 May 2007. Available online 10 May 2007. We would like to express our gratitude to Pat Leahy at Kerckhoff Marine Lab for his excellent care of the sea urchins. We would also like to thank Julie Hahn for generating the Gatae GFP BAC and for technical advice with BAC deletions. This work was supported by NIH grant HD-37105.

Additional details

Created:
August 22, 2023
Modified:
October 17, 2023