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Cloning and developmental expression of WSTF during Xenopus laevis embryogenesis.
Cus R
,
Maurus D
,
Kühl M
.
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The gene WSTF is deleted in the autosomal dominant hereditary disorder Williams-Beuren syndrome. This disorder is caused by a 1.3 megabase deletion in human chromosome 7, encompassing at least 17 genes. The WSTF protein contains a bromodomain, found predominantly in chromatin-associated proteins. Reported association of WSTF with chromatin remodeling factors and functional data support a role for WSTF during chromatin remodeling. Here, we report the cloning and developmental expression pattern of Xenopus laevis WSTF. Xenopus laevis WSTF is a protein with a predicted amino acid sequence of 1441 amino acids. Three discrete domains can be identified in the Xenopus laevis WSTF protein, a PHD finger, a DDT domain and a bromodomain. Alignment of Xenopus WSTF with the corresponding orthologues from Homo sapiens, Gallus gallus, Mus musculus and Danio rerio demonstrates an evolutionary conservation of WSTF amino acid sequence and domain organization. In situ hybridization reveals a dynamic expression profile during embryonic development. WSTF is expressed differentially in neural tissue, especially during neurulae stages in the eye, in neural crest cells and the brain.
Fig. 2. Expression of WSTF during Xenopus laevis embryonic development. (A) At stage 18 WSTF is expressed in the closing neural tube. Dashed line indicates plane of section in Fig. 3A. (B) At stage 20/21, expression of WSTF is very strong in the optic cup (arrow), in migrating neural crest cells (arrowhead) and the anteriorbrain. WSTF expression is also detectable in the neural tube, although to a lower extent. (C and D) WSTF expression at stage 24/25 persists strongly in the optic cup (arrow) and the migrating neural crest cells (arrowhead) and weaker in the neural tube. Dashed line indicates plane of section in Fig. 3B. (E) At stage 32, expression of WSTF in the forebrain has ceased whereas expression in the dorsal midbrain (tectum) and the mid/hindbrain-boundary (isthmus) is still bold (arrowhead). Pronounced WSTF expression is also detectable in the branchial arches (white arrowhead). Another tissue exhibiting WSTF expression is the embryonic tailbud. Dashed line indicates plane of section in Fig. 3C.
Fig. 3. Sections of embryos subjected to whole-mount in situ hybridization with a WSTF probe. (A) Horizontal section of an embryo stage 18. Expression is restricted to neural tissue. (B) Cross-bisected embryo stage 25. Expression in the neural tube is weakly detectable (arrowhead). (C) Horizontally bisected embryo stage 32. Cross section of the branchial arches reveals that staining is restricted to the interior of the branchial arches (arrowhead). Ectoderm and endoderm lining of the branchial arch are devoid of expression. (D) Cross section through head region of an embryo stage 27. WSTF is strongly expressed in the optic cup (arrow). WSTF expression is also traceable in mesenchym surrounding the stomodeum (arrowhead). (E) At stage 30 expression of WSTF in the eye has retracted to the ciliary marginal zone (arrow). (F) Blow-up of eye region of a different section indicating expression in the cilliary marginal zone.
