Domino S E, Hiraiwa N, Lowe J B
Department of Obstetrics and Gynecology, The University of Michigan Medical School, Ann Arbor, Michigan 48109-0650, USA.
Biochem J. 1997 Oct 1;327 ( Pt 1)(Pt 1):105-15. doi: 10.1042/bj3270105.
Terminal Fucalpha(1-2)Galbeta epitopes have been proposed to play significant roles in cell-cell interactions in development, cell adhesion, and malignant transformation. To begin to investigate the regulation and function of alpha(1-2)fucosylated epitopes in an animal model, we have isolated and characterized a mouse genomic DNA segment encoding a protein orthologous to the human H blood group locus alpha(1,2)fucosyltransferase (FUT1). This segment maintains an open reading frame encoding 376 amino acids sharing 75% sequence identity with the enzyme encoded by human FUT1, and 55% sequence identity with the enzyme encoded by the human Secretor blood group locus (FUT2). Expression of the open reading frame in COS-7 cells yields an alpha(1,2)fucosyltransferase activity with a Km of 7.6 mM for phenyl-beta-d-galactoside. Southern blotting and interspecific backcross analyses indicate that this murine locus represents a single copy sequence mapping to a novel locus 2.1 centimorgans from the Klk1 locus, in a region of homology between mouse chromosome 7 and the human FUT1 locus on the long arm of chromosome 19. Mouse FUT1 yields a 2.8 kb mRNA transcript identifiable in many organs, including thymus, lung, stomach, pancreas, small intestine, colon, uterus and epidiymis. Hybridization analyses in situ localize expression of FUT1 transcripts to thymic medullary and epididymal epithelial cells, implying that this gene determines the expression of cell surface Fucalpha(1-2)Galbeta epitopes in these tissues.
末端Fucα(1-2)Galβ表位已被认为在发育过程中的细胞间相互作用、细胞黏附及恶性转化中发挥重要作用。为了在动物模型中开始研究α(1-2)岩藻糖基化表位的调控及功能,我们分离并鉴定了一个小鼠基因组DNA片段,其编码一种与人H血型位点α(1,2)岩藻糖基转移酶(FUT1)直系同源的蛋白质。该片段维持一个开放阅读框,编码376个氨基酸,与人类FUT1编码的酶具有75%的序列同一性,与人类分泌型血型位点(FUT2)编码的酶具有55%的序列同一性。该开放阅读框在COS-7细胞中的表达产生一种α(1,2)岩藻糖基转移酶活性,对苯基-β-D-半乳糖苷的Km为7.6 mM。Southern印迹和种间回交分析表明,这个小鼠基因座代表一个单拷贝序列,定位于距Klk1基因座2.1厘摩的一个新基因座,位于小鼠7号染色体与人类19号染色体长臂上FUT1基因座之间的同源区域。小鼠FUT1产生一个2.8 kb的mRNA转录本,在许多器官中均可识别,包括胸腺、肺、胃、胰腺、小肠、结肠、子宫和附睾。原位杂交分析将FUT1转录本的表达定位到胸腺髓质和附睾上皮细胞,这意味着该基因决定了这些组织中细胞表面Fucα(1-2)Galβ表位的表达。