The cis-regulatory system of the tbrain gene: Alternative use of multiple modules to promote skeletogenic expression in the sea urchin embryo
Abstract
The genomic cis-regulatory systems controlling regulatory gene expression usually include multiple modules. The regulatory output of such systems at any given time depends on which module is directing the function of the basal transcription apparatus, and ultimately on the transcription factor inputs into that module. Here we examine regulation of the Strongylocentrotus purpuratus tbrain gene, a required activator of the skeletogenic specification state in the lineage descendant from the embryo micromeres. Alternate cis-regulatory modules were found to convey skeletogenic expression in reporter constructs. To determine their relative developmental functions in context, we made use of recombineered BAC constructs containing a GFP reporter and of derivatives from which specific modules had been deleted. The outputs of the various constructs were observed spatially by GFP fluorescence and quantitatively over time by QPCR. In the context of the complete genomic locus, early skeletogenic expression is controlled by an intron enhancer plus a proximal region containing a HesC site as predicted from network analysis. From ingression onward, however, a dedicated distal module utilizing positive Ets1/2 inputs contributes to definitive expression in the skeletogenic mesenchyme. This module also mediates a newly discovered negative Erg input which excludes non-skeletogenic mesodermal expression.
Additional Information
© 2009 Elsevier Inc. Received 27 March 2009; revised 24 July 2009; accepted 3 August 2009. Available online 11 August 2009. We would like to thank Prof. Andrew Murray and anonymous reviewers for critical reading and helpful suggestions. Research was supported by the Caltech SURF program, the Camilla Chandler Frost Fellowship, the U.S. Department of Defense NDSEG Fellowship Program, and NIH grants HD037105 and GM075089.Attached Files
Accepted Version - nihms-151364.pdf
Supplemental Material - applic1.pdf
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Additional details
- PMCID
- PMC2784196
- Eprint ID
- 16782
- Resolver ID
- CaltechAUTHORS:20091123-113132667
- Caltech SURF program
- Camilla Chandler Frost Fellowship
- U. D. Department of Defense NDSEG Fellowship
- NIH
- HD037105
- NIH
- GM075089
- Created
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2009-11-24Created from EPrint's datestamp field
- Updated
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2021-11-08Created from EPrint's last_modified field