CHARACTERIZATION OF CDNAS ENCODING HUMAN LEUKOSIALIN AND LOCALIZATION OF THE LEUKOSIALIN GENE TO CHROMOSOME-16
CHARACTERIZATION OF CDNAS ENCODING HUMAN LEUKOSIALIN AND LOCALIZATION OF THE LEUKOSIALIN GENE TO CHROMOSOME-16
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DOI：
10.1073/pnas.86.4.1328
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发表时间：
1989-02-01
影响因子：
11.1
通讯作者：
FRELINGER, JG
中科院分区：
文献类型：
--
作者：
PALLANT, A;ESKENAZI, A;FRELINGER, JG
我们描述了人类白细胞的一种主要唾液酸糖蛋白--人白涎素的编码基因克隆的分离和鉴定。白唾液素与唾液酸分子密切相关或相同，唾液酸分子参与T细胞增殖，其表达在Wiskott-Aldrich综合征(Wiskott-Aldrich综合征，一种X染色体连锁免疫缺陷疾病)中发生变化。用兔抗白唾液酸抗血清，从人外周血细胞构建的λgt11基因文库中克隆了一个cDNA克隆。该lambda.gt11克隆被用来分离与白唾液素的整个编码序列相对应的更长的cDNA克隆。DNA序列分析显示，在预测的成熟蛋白中有三个结构域。胞外区富含丝氨酸、苏氨酸和Pro，并包含四个连续的18个氨基酸重复序列。人白涎素分子的跨膜区和胞内区与大鼠W3/13分子高度同源。RNA凝胶印迹分析显示有两个多腺化物种，分别为2.3kb和8kb。Southern杂交分析表明，人类白涎素是一个单拷贝基因。对单染色体细胞杂交的分析表明，白唾液素基因不是X染色体连锁的，原位杂交表明白唾液素基因位于第16号染色体上。这些结果表明，该突变不是白唾液素结构基因的缺陷。
We describe the isolation and characterization of cDNA clones encoding human leukosialin, a major sialoglycoprotein of human leukocytes. Leukosialin is very closely related or identical to the sialophorin molecule, which is involved in T-cell proliferation and whose expression is altered in Wiskott-Aldrich syndrome (WAS), an X chromosome-linked immunodeficiency disease. Using a rabbit antiserum to leukosialin, a cDNA clone was isolated from a .lambda.gt11 cDNA library constructed from human peripheral blood cells. This .lambda.gt11 clone was used to isolate longer cDNA clones that correspond to the entire coding sequence of leukosialin. DNA sequence analysis reveals three domains in the predicted mature protein. The extracellular domain is enriched for Ser, Thr, and Pro and contains four contigous 18-amino acid repeats. The transmembrane and intracellular domains of the human leukosialin molecule are highly homologous to the rat W3/13 molecule. RNA gel blot analysis reveals two polyadenylylated species of 2.3 and 8 kilobases. Southern blot analysis suggests that human leukosialin is a single-copy gene. Analysis of monochromosomal cell hybrids indicates that the leukosialin gene is not X chromosome linked and in situ hybridization shows leukosialin is located on chromsome 16. These findings demonstrate that the primary mutation in WAS is not a defect in the structural gene for leukosialin.