Tbx6, Thylacine1, and E47 synergistically activate bowline expression in Xenopus somitogenesis

Hitachi, Keisuke; Kondow, Akiko; Danno, Hiroki; Inui, Masafumi; Uchiyama, Hideho; Asashima, Makoto

doi:10.1016/j.ydbio.2007.10.015

Cited by 20 publications

(21 citation statements)

References 42 publications

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“…The PCR product was subcloned into pGEM-T (Easy) vector (Promega) and named pGEM-6156. The pGEM-6156 construct was digested with MluI and inserted into MluI-digested pd2EGFP+s+N+M that had two restriction sites, NotI and MluI, added to pd2EGFP+s (Hitachi et al, 2008). The following constructs containing 5' sequence subdomain combinations, deletions or point mutations, were created from pd2EGFP-6156 (-6156) To generate pCS2-zVegT, pCS2-FoxH1-myc, pCS2-zVegT-myc, pCS2-Eomes-myc, pCS2-Xbra-myc, and pCS2-Tbx6-myc, the coding regions of zVegT, FoxH1, Eomes, Xbra, and Tbx6 were amplified by PCR then subcloned into pCS2+ or pCS2-MT (http://sitemaker.umich.edu/dlturner.vectors).…”

Section: Dna Constructs and Mrnamentioning

confidence: 99%

Zygotic VegT is required for Xenopus paraxial mesoderm formation and is regulated by Nodal signaling and Eomesodermin

Fukuda¹,

Takahashi²,

Haramoto³

et al. 2010

Int. J. Dev. Biol.

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The T-box gene VegT plays a crucial role during mesendoderm specification of the amphibian embryo. While the function of maternal VegT (mVegT) has been extensively investigated, little is known about the function and transcriptional regulation of zygotic VegT (zVegT). In the present study, we used comparative genomics and a knockdown experiment to demonstrate that zVegT is the orthologous gene of zebrafish Spadetail/Tbx16 and chick Tbx6L/Tbx6, and has an essential role in paraxial mesodermal formation. zVegT knockdown embryos show several defects in the patterning of trunk mesoderm, such as abnormal segmentation of somites, a reduction in muscle, and the formation of an abnormal mass of cells at the tail tip. We also identified the cis-regulatory elements of zVegT that are necessary and sufficient for mesodermspecific expression. These cis-regulatory elements are located in two separate upstream regions of zVegT, corresponding to the first intron of mVegT. The results of in vitro binding and functional assays indicate that Forkhead box H1 (FoxH1) and Eomesodermin (Eomes) are the trans-acting factors required for zVegT expression. Our results highlight the essential role of zVegT in organization of paraxial mesoderm, and reveal that zVegT is regulated by a coherent feedforward loop of Nodal signaling via Eomes.

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Section: Dna Constructs and Mrnamentioning

confidence: 99%

Zygotic VegT is required for Xenopus paraxial mesoderm formation and is regulated by Nodal signaling and Eomesodermin

Fukuda¹,

Takahashi²,

Haramoto³

et al. 2010

Int. J. Dev. Biol.

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“…We have shown previously that the expression of bowline is activated by the combined effects of three transcriptional factors, Tbx6, mespb, and E47 (Hitachi et al, 2008b). We examined whether the expression of ledgerline is also regulated by these three factors.…”

Section: Tbx6 Is Indispensable For the Expression Of Both Bowline Andmentioning

confidence: 99%

“…To produce the pGL4-Tbox construct, the Tbx1 binding site (5'-TTTCACACCTAGGTGTGAAA-3'), as described previously (Ataliotis et al, 2005), was tandemly inserted into the pGL4.23 vector (Promega). The pCS2-Bowline, pCS2-Grg4, pCS2-Tbx6-VP16, pCS2-Xotch∆E, pCS2-mespb (previously referred to as pCS2-Thy1), pCS2-E47, pCS2-mycTbx6, and pCS2-myc-Grg4 plasmids have been described previously (Hitachi et al, 2008b;Kondow et al, 2006;Kondow et al, 2007;Uchiyama et al, 2001). The pGL4.2Thy1 construct, which contains the 5'-flanking region of Xenopus mespb, has also been described previously .…”

Section: Plasmid Constructsmentioning

confidence: 99%

“…In addition, we reveal the transcription factors involved in the regulation of ledgerline expression. Tbx6, mespb, and E47 regulate bowline expression during Xenopus somitogenesis (Hitachi et al, 2008b). Overexpression of Tbx6, mespb, and E47 induced ectopic ledgerline expression, as well as bowline expression, in early-stage Xenopus embryos.…”

mentioning

confidence: 96%

“…for the bowline, mespb, and Tbx6 probes have been described (Hitachi et al, 2008b). The pCS2-Ledgerline, pBS-Tbx1, pBS-xRipply3, and pBSmRipply3 constructs were used as templates for the probes.…”

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confidence: 99%

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The Xenopus Bowline/Ripply family proteins negatively regulate the transcriptional activity of T-box transcription factors

et al. 2009

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Bowline, which is a member of the Xenopus Bowline/Ripply family of proteins, represses the transcription of somitogenesis-related genes before somite segmentation, which makes Bowline indispensable for somitogenesis. Although there are three bowline/Ripply family genes in each vertebrate species, it is not known whether the Bowline/Ripply family proteins share a common role in development. To elucidate their developmental roles, we examined the expression patterns and functions of the Xenopus Bowline/Ripply family proteins Bowline, Ledgerline, and a novel member of this protein family, xRipply3. We found that the expression patterns of bowline and ledgerline overlapped in the presomitic mesoderm (PSM), whereas ledgerline was additionally expressed in the newly formed somites. In addition, we isolated xRipply3, which is expressed in the pharyngeal region. Co-immunoprecipitation assays revealed that Ledgerline and xRipply3 interacted with T-box proteins and the transcriptional co-repressor Groucho/TLE. In luciferase assays, xRipply3 weakly suppressed the transcriptional activity of Tbx1, while Ledgerline strongly suppressed that of Tbx6. In line with the repressive role of Ledgerline, knockdown of Ledgerline resulted in enlargement of expression regions of the somitogenesis-related-genes mespb and Tbx6. Inhibition of histone deacetylase activity increased the expression of mespb, as seen in the Bowline and Ledgerline knockdown experiments. These results suggest that the Groucho-HDAC complex is required for the repressive activity of Bowline/Ripply family proteins during Xenopus somitogenesis. We conclude that although the Xenopus Bowline/Ripply family proteins Bowline, Ledgerline and xRipply3 are expressed differentially, they all act as negative regulators of T-box proteins.

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tbx6l and tbx16 are redundantly required for posterior paraxial mesoderm formation during zebrafish embryogenesis

Morrow

Maxwell

Hoshijima

et al. 2017

Developmental Dynamics

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Background T-box genes encode a large transcription factor family implicated in many aspects of development. We are focusing on two related zebrafish T-box genes, tbx6l and tbx16, that are expressed in highly overlapping patterns in embryonic paraxial mesoderm. tbx16 mutants are deficient in trunk, but not tail, somites; we explored whether presence of tail somites in tbx16 mutants was due to compensatory function provided by the tbx6l gene. Results We generated two zebrafish tbx6l mutant alleles. Loss of tbx6l has no apparent effect on embryonic development, nor does tbx6l loss enhance the phenotype of two other T-box gene mutants, ta or tbx6, or of the mesp family gene mutant, msgn1. In contrast, loss of tbx6l function dramatically enhances the paraxial mesoderm deficiency of tbx16 mutants. Conclusion These data demonstrate that tbx6l and tbx16 genes function redundantly to direct tail somite development. tbx6l single mutants develop normally because tbx16 fully compensates for loss of tbx6l function. However, tbx6l only partially compensates for loss of tbx16 function. These results resolve the question of why loss of function of tbx16 gene, which is expressed throughout the ventral and paraxial mesoderm, profoundly affects somite development in the trunk but not the tail.

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Tbx6, Thylacine1, and E47 synergistically activate bowline expression in Xenopus somitogenesis

Cited by 20 publications

References 42 publications

Zygotic VegT is required for Xenopus paraxial mesoderm formation and is regulated by Nodal signaling and Eomesodermin

Zygotic VegT is required for Xenopus paraxial mesoderm formation and is regulated by Nodal signaling and Eomesodermin

The Xenopus Bowline/Ripply family proteins negatively regulate the transcriptional activity of T-box transcription factors

tbx6l and tbx16 are redundantly required for posterior paraxial mesoderm formation during zebrafish embryogenesis

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