MOLECULAR BIOLOGY OF HUMAN ALPHA 1,3 FUCOSYLTRANSFERASE

这项工作的总体目标是确定监管机制 哺乳动物低聚糖分子的表达。岩藻糖化 寡糖被选为实验模型是因为它们的 岩藻糖键和同源岩藻糖基转移酶 它们的生物合成，在时间和空间上都有精确的表达 在哺乳动物分化过程中，并经常在 与恶变有关。这些属性表明 这些分子的重要功能；这是最近才发现的 通过研究表明，特定集合中的一个或多个成员 岩藻糖化低聚糖，由髓系细胞表达， 这些白细胞与内皮细胞白细胞黏附分子I的黏附 在发炎期间。 为了解决这些分子的表达调控问题，我们已经 分离的基因和cDNA编码三个不同的 α(1，3)岩藻糖基转移酶。这些工具可用于 研究转录、转录后和结构功能 决定不同类型和相对数量的考虑因素 由细胞和组织制成的α(1，3)岩藻糖化糖偶联物。 这项建议的具体目标是： 1.分析一个新的人类DNA序列的结构和功能 可能编码一个α(1.3)岩藻糖基转移酶。 2.分离克隆的髓系cDNAs 细胞表面唾液酸化Lewis x和vim-2决定簇的生物合成。 3.定义每个α(1，3)FT的生物合成和结构，使用 生物化学、分子遗传学和形态学方法。 4.探讨α(1，3)FT受体之间的关系 底物专一性和主要序列决定因素 通过创建和测试嵌合、突变的α(1，3)FTs α(1，3)FTs。 5.明确正常人α(1，3)FT基因的表达模式 纸巾。

The overall goals of this work are to determine mechanisms that regulate expression of mammalian oligosaccharide molecules. Fucosylated oligosaccharides were chosen as an experimental model because their fucose linkages, and the cognate fucosyltransferases responsible for their biosynthesis, are expressed with temporal and spatial precision during mammalian differentiation, and are frequently altered in association with malignant transformation. These properties suggested important functions for these molecules; this has been born out recently by studies demonstrating that one or more members of a specific set of fucosylated oligosaccharides, expressed by cells of the myeloid lineage, adhesion of these leukocytes to endothelial leukocyte adhesion molecule I during inflammation. To address the regulation of expression of such molecules, we have isolated genes and cDNAs that encode three distinct alpha(1,3)fucosyltransferases. These represent tools with which to examine transcriptional, post-transcriptional, and structure-function considerations that determine the types and relative amounts of distinct alpha(1,3)fucosylated glycoconjugates made by cells and tissues. The SPECIFIC AIMS of this proposal are: 1. To analyze the structure and function of a novel human DNA sequence that may encode an alpha(1.3)fucosyltransferase. 2. To isolate cloned, myeloid-lineage cDNAs that participate in the biosynthesis of cell surface sialyl Lewis x and VIM-2 determinants. 3. To define the biosynthesis and structure of each alpha(1,3)FT, using biochemical, molecular genetic, and morphologic approaches. 4. To investigate the relationship between alpha(1,3)FT acceptor substrate specificity and primary sequence determinants within alpha(1,3)FTs via the creation and testing of chimeric, mutant alpha(1,3)FTs. 5. To define expression patterns of alpha(1,3)FT genes in normal human tissues.