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Expression analysis of Runx3 and other Runx family members during Xenopus development.

ABSTRACT: Runx genes encode a family of proteins defined by the highly conserved runt DNA-binding domain. Studies in several organisms have shown that these transcription factors regulate multiple aspects of embryonic development and are responsible for the pathogenesis of several human diseases. Here we report the cloning and expression of Runx3 during Xenopus development and compare its expression pattern to other Runx family members, Runx1 and Runx2, and to Cbfbeta, the obligatory binding partner of Runx proteins. Using in situ hybridization in the whole embryo and on sections we show that Runx3 is co-expressed with Runx1 in the hematopoietic lineage and in Rohon-Beard sensory neurons. In contrast Runx3 and Runx2 are co-expressed in craniofacial cartilage elements. Runx3 shows also unique expression domains in a number of derivatives of the neurogenic placodes, including the ganglia of the anteroposterior and middle lateral line nerves, and ganglia of the trigeminal, glossopharyngeal, facial and vagal nerves. These observations suggest a critical role for Runx3 in the development of cranial sensory neurons, while in other tissues its co-expression with Runx1 or Runx2 may signify functional redundancy between these family members.

SUBMITTER: Park BY

PROVIDER: S-EPMC2896033 | biostudies-literature | 2010 Jun

REPOSITORIES: biostudies-literature

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Expression analysis of Runx3 and other Runx family members during Xenopus development.

Park Byung-Yong BY Saint-Jeannet Jean-Pierre JP

Gene expression patterns : GEP 20100428 4-5

Runx genes encode a family of proteins defined by the highly conserved runt DNA-binding domain. Studies in several organisms have shown that these transcription factors regulate multiple aspects of embryonic development and are responsible for the pathogenesis of several human diseases. Here we report the cloning and expression of Runx3 during Xenopus development and compare its expression pattern to other Runx family members, Runx1 and Runx2, and to Cbfbeta, the obligatory binding partner of Ru ...[more]

PMID: 20433948

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Project description:Runx transcription factors play pivotal roles in the development of hematopoietic cells, including T cells. However, the Runx-target genes in early stages of thymic T cell development have been only partially elucidated. Moreover, the relationships of different Runx paralogs, whether they compete or collaborate with each other, are unknown. We have performed CRISPR-Cas9 mediated acute deletion of Runx1 and Runx3, alone and in combination, to define the functions of Runx factors and their target genes. We focused on two distinct stages of early T cell development; before lineage commitment (Phase1) and after commitment (Phase2). Our data suggest that Runx1 and Runx3 are functionally redundant in Phase1, hence the absence of one factor was compensated by the other factor. In Phase2, Runx1 becomes a major contributor, while low levels of Runx3 still strengthened the Runx1-mediated target gene regulation and showed an additive effect. Of importance, we found that Runx1 and Runx3 upregulate T cell programs and inhibit alternative lineage genes. In order to capture the developmental status of Runx-perturbed cells, we performed supervised principal component analysis (PCA) using a fixed frame of PC loadings from single cell RNA-seq analysis of normal in vivo-derived thymocytes. Unlike the reference in vitro and in vivo pro-T cell population or sgControl introduced cells, Runx knockout cells either failed to progress fully (Phase1) or deflected from the normal developmental trajectory (Phase2), suggesting essential roles of Runx factors in early stages of T development. In addition, we analyzed the correlation between stage-specific genomic occupancy of Runx1 and Runx3 (using previously published data for Runx1, GSE103953) and Runx-mediated gene regulation responses at each stage. Interestingly, genes with dynamic, stage-sensitive expression showed a very significant association with phase-specific genomic interactions of Runx factors, and an especially strong correlation with Runx activated target genes. In summary, our data suggest a pivotal role for Runx transcription factors in thymic T cell development by imposing T lineage specification in Phase1 and instructing the lineage-committed progenitor cells to progress through a normal T developmental trajectory.

Dataset Information

Expression analysis of Runx3 and other Runx family members during Xenopus development.

Publications

Expression analysis of Runx3 and other Runx family members during Xenopus development.

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