An extracellular matrix response element in the promoter of the LpS1 genes of the sea urchin Lytechinus pictus

Christopher A. Seid, Ravi K. Ramachandran, Jenny M. George, Venkatesh Govindarajan, Maria F. González-Rimbau, Constantin N. Flytzanis, Craig R. Tomlinson

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Abstract

The extracellular matrix (ECM) has been shown to play an important role in development and tissue-specific gene expression, yet the mechanism by which genes receive signals from the ECM is poorly understood. The aboral ectoderm-specific LpS1-α and -β genes of Lytechinus pictus, members of the Spec gene family, provide an excellent model system to study ECM-mediated gene regulation. Disruption of the ECM by preventing collagen deposition using the lathrytic agent β-aminopropionitrile (BAPN) inhibits LpS1 gene transcription. LpS1 transcription resumes after removal of BAPN and subsequent collagen reformation. Using a chloramphenicol acetyltransferase (CAT) reporter gene assay, we show that a 125 bp region of the LpS1-β promoter from -108 to +17 contains an ECM response element (ECM RE). Insertion of the 125 bp region into the promoter of the metallothionein gene of L. pictus, a gene unaffected by ECM disruption, caused the fused promoter to become ECM dependent. As with the endogenous LpS1 genes, CAT activity directed by the fused LpS1-β promoter resumed in embryos recovered from ECM disruption. A mutation in a cis-acting element called the proximal G-string, which lies in the 125 bp region, caused CAT activity levels in ECM-disrupted embryos to equal that of the wild-type LpS1-β promoter in ECM-intact embryos. These results suggest that the intact ECM normally transmits signals to inhibit repressor activity at the proximal G-string in aboral ectoderm cells. Consistent with these results were our findings which showed that in addition to expression in the aboral ectoderm, the proximal G-string mutation caused expression of the CAT gene in oral ectoderm cells. These studies suggested that the proximal G-string serves as a binding site for negative regulation of the LpS1 genes in oral ectoderm during development. We also examined trans-acting factors binding the proximal G-string following ECM disruption. Band shift gels revealed a predominant set of slower migrating nuclear proteins from ECM-disrupted embryos which bound the proximal G-string. This work suggested that ECM disruption initiates signaling that induces a repressor to bind the ECM RE and/or modifies ECM RE binding proteins, which in turn represses LpS1 gene activity.

Original languageEnglish (US)
Pages (from-to)3175-3182
Number of pages8
JournalNucleic Acids Research
Volume25
Issue number15
DOIs
StatePublished - Aug 1 1997

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All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Seid, C. A., Ramachandran, R. K., George, J. M., Govindarajan, V., González-Rimbau, M. F., Flytzanis, C. N., & Tomlinson, C. R. (1997). An extracellular matrix response element in the promoter of the LpS1 genes of the sea urchin Lytechinus pictus. Nucleic Acids Research, 25(15), 3175-3182. https://doi.org/10.1093/nar/25.15.3175