YY1 regulates the neural crest-associated slug gene in Xenopus laevis.
slug gene expression is associated with the specification and migration of neural crest cells in the African clawed frog Xenopus laevis. We provide evidence that the protein Ying-Yang 1 (YY1) regulates the slug gene expression both indirectly and directly, via a YY1 cis-element in the slug promoter, during Xenopus development. The ability of the YY1 to bind this YY1 cis-element was confirmed by electromobility shift assays and reporter assays. YY1 was detected in the nuclei of ectodermal cells contemporaneously with the process of neural crest specification. The injection of anti-YY1 morpholino, which targeted both YY1alpha and YY1beta gene products, depleted YY1 expression below 20% and was lethal at gastrulation. Sublethal depletion of YY1 reduced the length of the anterior-posterior axis and severely inhibited the expression of the neural marker Nrp1 and of the slug gene. Overexpression of YY1 or mutation of the YY1 cis-element reduced the restricted spatial expression of the slug reporter gene in the neural ectoderm border and provoked its expression in the nonneural ectoderm. Chromatin immunoprecipitation indicated that endogenous YY1 interacts directly with the YY1 cis-element of the endogenous slug gene and with the slug gene reporter sequence injected into embryos. The results suggest that YY1 is essential for Xenopus development; is necessary for neural ectoderm differentiation, a prerequisite for neural crest specification; and restricts which cells can form neural crest mesenchyme through directly blocking slug gene activity.
Morgan
Matthew J
MJ
Institut Curie-CNRS UMR144, Morphogénèse Cellulaire et Progression Tumorale, 26 Rue d'Ulm, 75248 Paris Cedex 05, France. mattew.morgan@curie.fr
Woltering
Joost M
JM
In der Rieden
Paul M J
PM
Durston
Antony J
AJ
Thiery
Jean Paul
JP
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D050981:Erythroid-Specific DNA-Binding Factors; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006720:Homozygote; D020287:In Situ Nick-End Labeling; D008648:Mesoderm / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009154:Mutation; D009432:Neural Crest / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012313:RNA / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D000071250:Snail Family Transcription Factors; D013997:Time Factors; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic; D029867:Xenopus Proteins; D014982:Xenopus laevis; D050986:YY1 Transcription Factor
D017931:DNA Primers; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D016376:Oligonucleotides, Antisense; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D029867:Xenopus Proteins; D050986:YY1 Transcription Factor; C494701:YY1 protein, Xenopus; D049452:Green Fluorescent Proteins; D012313:RNA