XIC Is Required for Siamois Activity and Dorsoanterior Development

Abstract

Siamois is the transcriptional mediator of the dorsal Wnt signaling pathway and is necessary for formation of the Spemann organizer and dorsoanterior development in Xenopus. We have determined that XIC, a Xenopus I-mfa domain protein that regulates Tcf3 binding, is required for dorsoaxial development and specifically for Siamois activity in establishing the dorsal organizer. In loss-of-function studies, we found that embryos injected with a morpholino to XIC mRNA (XIC morphpolino) are missing head structures, neural tube, notochord, and paraxial mesoderm as well as NCAM and XMyoD expression. Although Siamois, Twin, and Xnr3 expression is normal in morpholino-injected embryos, levels of downstream organizer factors, including goosecoid, Xnot, Cerberus, and chordin, are severely reduced. Ectopic axis formation induced by Siamois is repressed by injection of the XIC morpholino and further repressed by coinjection of β-catenin or a constitutively active Tcf3/HMG/G4A fusion. Activation of reporters driven by the Siamois-responsive proximal element of the goosecoid promoter is inhibited in the presence of the morpholino and can be rescued by murine I-mfa and by a dominant-negative Tcf3. The data indicate a role for XIC in limiting Tcf3-dependent repression of Siamois activities that are required for goosecoid transcription and for dorsal organizer formation.

ACKNOWLEDGMENTS

We thank R. T. Moon, K. Cho, and D. Kimelman for plasmids and members of the Tapscott laboratory for support and discussions.

This work was supported by NIAMS AR45113 to S.J.T.