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XB-ART-60641
Mol Cells 2024 Mar 23;474:100058. doi: 10.1016/j.mocell.2024.100058.
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Cdx1 and Gsc distinctly regulate the transcription of BMP4 target gene ventx3.2 by directly binding to the proximal promoter region in Xenopus gastrulae.

Goutam RS , Kumar V , Lee U , Kim J .


Abstract
A comprehensive regulatory network of transcription factors controls the dorsoventral patterning of the body axis in developing vertebrate embryos. Bone morphogenetic protein signaling is essential for activating the Ventx family of homeodomain transcription factors, which regulates embryonic patterning and germ layer identity during Xenopus gastrulation. Although Ventx1.1 and Ventx2.1 of the Xenopus Ventx family have been extensively investigated, Ventx3.2 remains largely understudied. Therefore, this study aimed to investigate the transcriptional regulation of ventx3.2 during the embryonic development of Xenopus. We used goosecoid (Gsc) genome-wide ChIP-sequencing data to isolate and replicate the promoter region of ventx3.2. Serial deletion and site-directed mutagenesis were used to identify the cis-acting elements for Gsc and caudal type homeobox 1 (Cdx1) within the ventx3.2 promoter. Cdx1 and Gsc differentially regulated ventx3.2 transcription in this study. Additionally, positive cis-acting and negative response elements were observed for Cdx1 and Gsc, respectively, within the 5' flanking region of the ventx3.2 promoter. This result was corroborated by mapping the active Cdx1 response element (CRE) and Gsc response element (GRE). Moreover, a point mutation within the CRE and GRE completely abolished the activator and repressive activities of Cdx1 and Gsc, respectively. Furthermore, chromatin immunoprecipitation-polymerase chain reaction confirmed the direct binding of Cdx1 and Gsc to the CRE and GRE, respectively. Inhibition of Cdx1 and Gsc activities at their respective functional regions, namely, the ventral (VMZ) and dorsal (DMZ) marginal zones, reversed their effects on ventx3.2 transcription. These results indicate that Cdx1 and Gsc modulate ventx3.2 transcription in the VMZ and DMZ by directly binding to the promoter region during Xenopus gastrulation.

PubMed ID: 38522664
PMC ID: PMC11031840
Article link: Mol Cells


Species referenced: Xenopus laevis
Genes referenced: bmp4 cdx1 gsc myc smad1 tbxt ventx1 ventx1.2 ventx2 ventx2.2 ventx3 ventx3.2
GO keywords: gastrulation [+]
Morpholinos: cdx1 MO4 gsc MO1

Phenotypes: Xbo Wt + gsc MO + cdx1 MO (VMZ) (Supp. Fig 4 E) [+]

Article Images: [+] show captions