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Genes and Development
Vol. 11, No. 17,
pp. 2176-2190,
September 1, 1997
1 Center for Developmental Biology, Department of
Zoology, University of Texas at Austin, Austin, Texas 78712 USA;
2 Wellcome/Cancer Research Campaign (CRC)
Institute of Cancer and Developmental Biology, Cambridge CB2 1QR, UK
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.
[Key Words: quaking; Xqua; KH domain; RNA binding; notochord; Xenopus]
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