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Characterization of Cer-1 cis-regulatory region during early Xenopus development

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Abstract

Cerberus-related molecules are well-known Wnt, Nodal, and BMP inhibitors that have been implicated in different processes including anterior–posterior patterning and left–right asymmetry. In both mouse and frog, two Cerberus-related genes have been isolated, mCer-1 and mCer-2, and Xcer and Xcoco, respectively. Until now, little is known about the mechanisms involved in their transcriptional regulation. Here, we report a heterologous analysis of the mouse Cerberus-1 gene upstream regulatory regions, responsible for its expression in the visceral endodermal cells. Our analysis showed that the consensus sequences for a TATA, CAAT, or GC boxes were absent but a TGTGG sequence was present at position −172 to −168 bp, relative to the ATG. Using a series of deletion constructs and transient expression in Xenopus embryos, we found that a fragment of 1.4 kb of Cer-1 promoter sequence could reproduce the endogenous expression pattern of Xenopus cerberus. A 0.7-kb mcer-1 upstream region was able to drive reporter expression to the involuting mesendodermal cells, while further deletions abolished reporter gene expression. Our results suggest that although no sequence similarity was found between mouse and Xenopus cerberus cis-regulatory regions, the signaling cascades regulating cerberus expression, during gastrulation, is conserved.

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Abbreviations

AVE:

Anterior visceral endoderm

ADME:

Anterior dorsal mesendoderm

dkk-1:

dickkopf-1

Xcer :

Xenopus Cerberus

mcer-1:

Mouse cerberus-like

IDME:

Involuting mesendoderm

ECR:

Evolutionary conserved sequence

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Acknowledgments

We thank Dr. A.T. Tavares for the cloning of the Xenopus Cerberus genomic region and A.T. Tavares, S. Marques, and R. Swain for critically reading of this manuscript. A. C. Silva and M. Filipe are recipients of F.C.T. PhD fellowships. This work was supported by research grants from IBB/CBME, LA, F.C.T., and IGC/Fundação Calouste Gulbenkian to J. A. Belo.

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Authors and Affiliations

  1. Regenerative Medicine Program, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, Faro, Portugal

    José António Belo

  2. IBB-Institute for Biotechnology and Bioengineering, Centro de Biomedicina Molecular e Estrutural, Universidade do Algarve, Campus de Gambelas, 8005-135, Faro, Portugal

    Ana Cristina Silva & José António Belo

  3. Instituto Gulbenkian de Ciência, Oeiras, Portugal

    Ana Cristina Silva, Mário Filipe & José António Belo

  4. Institute of Human Genetics, University of Heidelberg, Heidelberg, Germany

    Herbert Steinbeisser

  5. IBB-Institute for Biotechnology and Bioengineering, Dep. Ciências Biomédicas e Medicina, Edf. 8, Campus de Gambelas, Universidade do Algarve, 8005-139, Faro, Portugal

    José António Belo

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  1. Ana Cristina Silva
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  2. Mário Filipe
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  3. Herbert Steinbeisser
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  4. José António Belo
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Correspondence to José António Belo.

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Silva, A.C., Filipe, M., Steinbeisser, H. et al. Characterization of Cer-1 cis-regulatory region during early Xenopus development. Dev Genes Evol 221, 29–41 (2011). https://doi.org/10.1007/s00427-011-0357-5

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