Molecular cloning of a fourth member of a human alpha (1,3)fucosyltransferase gene family. Multiple homologous sequences that determine expression of the Lewis x, sialyl Lewis x, and difucosyl sialyl Lewis x epitopes.
Molecular cloning of a fourth member of a human alpha (1,3)fucosyltransferase gene family. Multiple homologous sequences that determine expression of the Lewis x, sialyl Lewis x, and difucosyl sialyl Lewis x epitopes.
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DOI：
10.1016/s0021-9258(18)35803-4
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发表时间：
1992-12
期刊：
影响因子：
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通讯作者：
Brent W. Weston;Peter L. Smith;R. Kelly;J. B. Lowe;J. B. Lowe
中科院分区：
文献类型：
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作者：
Brent W. Weston;Peter L. Smith;R. Kelly;J. B. Lowe;J. B. Lowe
我们和其他人以前已经描述了三个人α（1，3）岩藻糖基转移酶基因的分离，形成了一个新生的糖基转移酶基因家族的基础。我们现在报告的第四个同源的人α（1，3）岩藻糖基转移酶基因的分子克隆和表达。当转染到哺乳动物细胞中时，该岩藻糖基转移酶基因能够指导刘易斯x（Gal β 1->4[Fuc α 1->3]GlcNAc），唾液酸刘易斯x（NeuNAc α 2->3Gal β 1->4 [Fuc α 1->3]GlcNAc），和二岩藻糖基唾液酸刘易斯x（NeuNAc α 2->3Gal β 1->4[Fuc α 1->3]GlcNAc β 1-> 3Gal β 1->4[Fuc α 1->3]GlcNAc）表位。该酶与Fuc-TIII共享85%的氨基酸序列同一性，与Fuc-TV共享89%的氨基酸序列同一性，但在其受体底物要求方面存在显著差异。聚合酶链反应分析表明，该基因是同线的19号染色体上的Fuc-TIII和Fuc-TV。人基因组DNA的Southern印迹分析表明，这四个α（1，3）岩藻糖基转移酶基因占所有的DNA序列，在低严格性与Fuc-TIII催化结构域交叉杂交。使用类似的方法，来自Fuc-TIV的催化结构域探针鉴定了一类新的DNA片段，其不与染色体19岩藻糖基转移酶探针交叉杂交。这些结果扩展了人α（1，3）岩藻糖基转移酶基因家族的分子定义，并提供了检测岩藻糖基转移酶基因表达的工具。
We and others have previously described the isolation of three human alpha (1,3)fucosyltransferase genes which form the basis of a nascent glycosyltransferase gene family. We now report the molecular cloning and expression of a fourth homologous human alpha (1,3)fucosyltransferase gene. When transfected into mammalian cells, this fucosyltransferase gene is capable of directing expression of the Lewis x (Gal beta 1–>4[Fuc alpha 1–>3]GlcNAc), sialyl Lewis x (NeuNAc alpha 2–>3Gal beta 1–>4 [Fuc alpha 1–>3]GlcNAc), and difucosyl sialyl Lewis x (NeuNAc alpha 2–>3Gal beta 1–>4[Fuc alpha 1–>3]GlcNAc beta 1–>3 Gal beta 1–>4[Fuc alpha 1–>3]GlcNAc) epitopes. The enzyme shares 85% amino acid sequence identity with Fuc-TIII and 89% identity with Fuc-TV but differs substantially in its acceptor substrate requirements. Polymerase chain reaction analyses demonstrate that the gene is syntenic to Fuc-TIII and Fuc-TV on chromosome 19. Southern blot analyses of human genomic DNA demonstrate that these four alpha (1,3)fucosyltransferase genes account for all DNA sequences that cross-hybridize at low stringency with the Fuc-TIII catalytic domain. Using similar methods, a catalytic domain probe from Fuc-TIV identifies a new class of DNA fragments which do not cross-hybridize with the chromosome 19 fucosyltransferase probes. These results extend the molecular definition of a family of human alpha (1,3)fucosyltransferase genes and provide tools for examining fucosyltransferase gene expression.