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

Pei Yun Lee, Jongmin Nam, Eric H. Davidson

Research output: Contribution to journalArticle

28 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)434-445
Number of pages12
JournalDevelopmental biology
Volume307
Issue number2
DOIs
StatePublished - Jul 15 2007
Externally publishedYes

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Embryonic Structures
Gastrula
Genes
Lytechinus
Strongylocentrotus purpuratus
Blastula
Endoderm
Mesoderm
Genetic Recombination
Sequence Analysis
Vertebrates
Exons
Gene Expression

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology
  • Developmental Biology

Cite this

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title = "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.",
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Exclusive developmental functions of gatae cis-regulatory modules in the Strongylocentrorus purpuratus embryo. / Lee, Pei Yun; Nam, Jongmin; Davidson, Eric H.

In: Developmental biology, Vol. 307, No. 2, 15.07.2007, p. 434-445.

Research output: Contribution to journalArticle

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