The KH domain protein encoded by quaking functions as a dimer and is essential for notochord development in Xenopus embryos

Aaron M. Zorn, Paul A. Krieg

Research output: Contribution to journalArticlepeer-review

73 Scopus citations

Abstract

Mutations in the mouse indicate that quaking gene function is essential for both embryogenesis and for development of the nervous system. Recent isolation of the mouse quaking gene identified a putative RNA-binding protein containing a single KH domain. We have previously isolated the Xenopus homolog of quaking, Xqua, and shown that the sequence is highly conserved through evolution. Here, we report experimental data on the biochemical function of the quaking protein and its role during development. We demonstrate that the quaking protein expressed during early embryogenesis, pXqua357, can bind RNA in vitro, and we have mapped the regions of the protein that are essential for RNA binding. We present evidence that pXqua can form homodimers and that dimerization may be required for RNA binding. Oocyte injection experiments show that pXqua357 is located in both the nucleus and cytoplasm. In the Xenopus embryo, Xqua is first expressed during gastrulation in the organizer region and its derivative, the notochord. In later stage embryos, Xqua is expressed in a number of mesodermal and neural tissues. We demonstrate that disruption of normal Xqua function, by overexpression of a dominant inhibitory form of the protein, blocks notochord differentiation. Xqua function appears to be required for the accumulation of important mRNAs such as Xnot, Xbra, and gsc. These results indicate an essential role for the quaking RNA-binding protein during early vertebrate embryogenesis.

Original languageEnglish (US)
Pages (from-to)2176-2190
Number of pages15
JournalGenes and Development
Volume11
Issue number17
DOIs
StatePublished - Sep 1 1997

Keywords

  • KH domain
  • Notochord
  • Quaking
  • RNA binding
  • Xenopus
  • Xqua

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

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