HUMAN PLASMINOGEN ACTIVATOR-INHIBITOR PAI-1 GENE

纤维蛋白溶解在某种程度上是由组织类型之间的平衡调节的。 纤溶酶原激活剂（tPA）及其特异性、快速作用 纤溶酶原激活物抑制剂（派-1）。 派-1合成 发生在许多组织中，包括血管内皮 细胞，并通过一个复杂的过程进行调节，涉及多个 因素 派-1的异常血浆水平与 与各种血栓栓塞性疾病的研究。 本提案中概述的目的是确定 人派-1基因，表征派-1基因的调控 在分子水平上表达，研究 派-1的结构和功能之间的关系，并开始研究PAI-1的结构和功能。 派-1遗传异常的分子遗传学基础。 在 初步研究，全长人派-1 cDNA已被 从λ gt 11 cDNA文库中分离， 序列确定。 人派-1基因已被定位 7号染色体 派-1序列显示出广泛的同源性 丝氨酸蛋白酶抑制剂（SERPIN） 超基因家族 派-1基因调控的初步研究 显示派-1 mRNA水平降低约10倍 在肝素和内皮细胞生长因子的存在下。 如果 这种肝素作用也发生在体内，这可能会显著增加 通过增加tPA活性， 随后纤维蛋白溶解增加。 在这份提案中，这些 观察结果将在分子水平上进一步表征 通过定量mRNA研究和引入重组 载体来评估特定调控序列的作用。 的 人派-1基因的详细结构将通过以下方法确定： 从重组质粒中分离派-1特异性克隆研究 人λ噬菌体基因组DNA文库。 比较 推导的外显子/内含子结构和启动子序列与 其他基因，特别是SERPIN基因家族， 协调控制相关基因的主要影响， 基因家族的进化 一个派-1基因限制性酶切长度 多态性（RFLP）已确定在初步研究 并将寻求更多的人。 这些工具最终可以 遗传连锁分析在遗传病家系中的应用 血栓栓塞性疾病

Fibrinolysis is regulated, in part, by a balance between tissue-type plasminogen activator (tPA) and its specific, rapidly acting inhibitor, plasminogen activator inhibitor (PAI-1). PAI-1 synthesis occurs in a number of tissues including the vascular endothelial cell and is regulated via a complex process involving multiple factors. Abnormal plasma levels of PAI-1 have been associated with a variety of thromboembolic disorders in man. The studies outlined in this proposal aim to determine the structure of the human PAI-1 gene, characterize the regulation of PAI-1 gene expression at the molecular level, investigate the relationship between PAI-1 structure and function, and begin to study the molecular genetic basis of hereditary abnormalities in PAI-1. In preliminary studies, full-length human PAI-1 cDNA has been isolated from a lambda gt11 cDNA library and its complete sequence determined. The human PAI-1 gene has been localized to chromosome 7. The PAI-1 sequence shows extensive homology to other members of the serine protease inhibitor (SERPIN) supergene family. Preliminary studies of PAI-1 gene regulation show PAI-1 mRNA levels to be decreased approximately 10-fold in the presence of heparin and endothelial cell growth factor. If this heparin affect also occurs in vivo, this could add significantly to heparin's clinical affect by increasing tPA activity with a subsequent increase in fibrinolysis. In this proposal, these observations will be further characterized at the molecular level by quantitative mRNA studies and introduction of recombinant vectors to assess the role of specific regulatory sequences. The detailed structure of the human PAI-1 gene will be determined by the study of PAI-1 specific clones isolated from a recombinant human lambda phage genomic DNA library. Comparison of the deduced exon/intron structure and promotor sequences to those of other genes, particularly the SERPIN gene family, may have major implications for coordinate control of related genes and the evolution of gene families. One PAI-1 gene restriction length polymorphism (RFLP) has been identified in preliminary studies and additional ones will be sought. These tools can eventually be applied to genetic linkage analysis in families with hereditary thromboembolic disease.