MOLECULAR-CLONING OF THE HUMAN PROTOONCOGENE WNT-5A AND MAPPING OF THE GENE (WNT5A) TO CHROMOSOME 3P14-P21
MOLECULAR-CLONING OF THE HUMAN PROTOONCOGENE WNT-5A AND MAPPING OF THE GENE (WNT5A) TO CHROMOSOME 3P14-P21
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DOI：
10.1006/geno.1993.1463
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发表时间：
1993-11-01
期刊：
影响因子：
4.4
通讯作者：
IOZZO, RV
中科院分区：
文献类型：
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作者：
CLARK, CC;COHEN, I;IOZZO, RV
高度保守的Wnt基因属于一个广泛分布的推定信号分子家族，其不仅涉及胚胎发生和细胞谱系分化过程中正常模式形成的调节，而且还涉及致癌事件。所有已知的脊椎动物Wnt基因编码38- 43-kDa富含半胱氨酸的推定糖蛋白，其具有分泌生长因子的典型特征：疏水信号序列，保守的天冬酰胺连接的寡糖共有序列，和22个保守的半胱氨酸残基，其相对间隔保持不变。在这项研究中，我们报告了几个重叠的cDNA编码约4.1 kb的人类同源的Wnt-5A的克隆和测序。成熟蛋白含有343个残基（Mr为38，000，不包括任何翻译后修饰），与已报道的其它Wnt-5A蛋白序列同源性>93%（与小鼠Wnt-5A同源性>99%）。这种蛋白质保持了某些特征-疏水信号序列，Wnt-1家族的“签名序列”（CKCHGvSGSC），和一些其他保守的氨基酸残基：24个半胱氨酸残基，4个天冬酰胺连接的寡糖的共识序列，和酪氨酸硫酸化位点-已发现在所有其他Wnt-5A蛋白。逆转录酶PCR分析的RNA从各种人类胚胎，新生儿，和成人细胞和/或组织显示，humanWnt-5A的表达只在新生儿的心脏和肺。然而，3′-非翻译区含有大量富含AT的基序可能与mRNA的快速降解有关。最后，利用Southern杂交、PCR扩增和原位杂交相结合的方法，将人Wnt-5A（WNT 5A）基因定位于染色体3 p14-p21。
The highly conservedWntgenes belong to a widely distributed family of presumptive signaling molecules that have been implicated not only in the regulation of normal pattern formation during embryogenesis and differentiation of cell lineages, but also in oncogenic events. All of the known vertebrateWntgenes encode for 38-to 43-kDa cysteine-rich putative glycoproteins, which have features typical of secreted growth factors: a hydrophobic signal sequence, a conserved asparagine-linked oligosaccharide consensus sequence, and 22 conserved cysteine residues whose relative spacing is maintained. In this study, we report the cloning and sequencing of several overlapping cDNAs encoding ∼4.1 kb of the human homologue ofWnt-5A. The mature protein contained 343 residues (Mr∼ 38,000 excluding any post-translational modifications) with a >93% homology to the reported sequences of otherWnt-5A proteins (>99% homologous to mouseWnt-5A). This protein maintained certain features—a hydrophobic signal sequence, theWnt-1 family "signature sequence" (CKCHGvSGSC), and a number of other conserved amino acid residues: 24 cysteine residues, 4 asparagine-linked oligosaccharide consensus sequences, and a tyrosine sulfation site—that have been found in all otherWnt-5A proteins. Reverse transcriptase PCR analysis of RNA from a variety of human embryonic, neonatal, and adult cells and/or tissues showed that humanWnt-5A expression was detected only in neonatal heart and lung. It may be relevant, however, that the 3′-untranslated region contained numerous AT-rich motifs that could be involved in the rapid degradation of mRNA. Finally, using a combination of Southern blotting, PCR amplification, andin situhybridization, the humanWnt-5A (WNT5A) gene was mapped to chromosome 3p14-p21.