An interferon regulatory factor-like binding element restricts Xmyf-5 expression in the posterior somites during Xenopus myogenesis

AbstractThe expression of myf-5, a key component of myogenic regulatory genes, declines progressively in mature somitic cells during vertebrate myogenesis. Little is known about how this down-regulation takes place. Here we provide evidence that an interferon regulatory factor binding element (IRF element) within the Xenopus myf-5 promoter is responsible for the elimination of myf-5 transcription in mature somitic mesoderm of Xenopus embryos. We show that this IRF element mediates the down-regulation of Xmyf-5 transcription in gastrula embryos, and can specifically interact with nuclear proteins of early neurula. Moreover, deletion of this IRF element results in the anterior expansion of reporter gene transcripts within somitic mesoderm in transgenic embryos. Our results, therefore, provide insight into how the negative control of Xmyf-5 expression takes place

Conserved Spätzle/Toll signaling in dorsoventral patterning of Xenopus embryos

The Spätzle/Toll signaling pathway controls ventral axis formation in Drosophila by generating a gradient of nuclear Dorsal protein. Dorsal controls the downstream regulators dpp and sog, whose patterning functions are conserved between insects and vertebrates. Although there is no experimental evidence that the upstream events are conserved as well, we set out to ask if a vertebrate embryo can respond to maternal components of the fly Dorsal pathway. Here we demonstrate a dorsalizing activity for the heterologous Easter, Spätzle and Toll proteins in UV-ventralized Xenopus embryos, which is inhibited by a co-injected dominant Cactus variant. We conclude that the Dorsal signaling pathway is a component of the conserved dorsoventral (d/v) patterning system in bilateria

CHD4/Mi-2β activity is required for the positioning of the mesoderm/neuroectoderm boundary in Xenopus

Experiments in Xenopus have illustrated the importance of extracellular morphogens for embryonic gene regulation in vertebrates. Much less is known about how induction leads to the correct positioning of boundaries; for example, between germ layers. Here we report that the neuroectoderm/mesoderm boundary is controlled by the chromatin remodeling ATPase CHD4/Mi-2β. Gain and loss of CHD4 function experiments shifted this boundary along the animal–vegetal axis at gastrulation, leading to excess mesoderm formation at the expense of neuroectoderm, or vice versa. This phenotype results from specific alterations in gene transcription, notably of the neural-promoting gene Sip1 and the mesodermal regulatory gene Xbra. We show that CHD4 suppresses Sip1 transcription by direct binding to the 5′ end of the Sip1 gene body. Furthermore, we demonstrate that CHD4 and Sip1 expression levels determine the “ON” threshold for Nodal-dependent but not for eFGF-dependent induction of Xbra transcription. The CHD4/Sip1 epistasis thus constitutes a regulatory module, which balances mesoderm and neuroectoderm formation