DEGRADATION OF E12/E47 IN SMOOTH MUSCLE CELLS

描述 （改编自申请人摘要） 血管平滑肌细胞（VSMC）是闭塞性血管炎的主要成分。 血管疾病，这是发达国家死亡的主要原因， 国家 作为一名预防心脏病专家，候选人的主要重点是 已经确定和治疗动脉粥样硬化的根本原因， 患有心血管疾病的患者。 作为一名调查员，他 旨在进一步了解这种疾病。 基本 螺旋-环-螺旋蛋白E12和E47促进细胞分化， 延缓细胞增殖。 它们在VSMC中表达。 申请人 克隆了一种新的泛素结合酶UBC-E2 A， E12和E47。 泛素-蛋白酶体系统可以显著改变 细胞表型通过调节反式作用因子的降解。 UBC-E2 A通过靶向E12和E47进行周转，可以促进VSMC 增殖和去分化。 本项目将以 VSMC中E12和E47蛋白降解。 首先，他将绘制UBC-E2 A 结合位点的E2 A蛋白，那么他将确定E2 A的作用 蛋白质磷酸化对其降解的影响。 这一分析将使 设计与UBC-E2 A竞争结合E2 A蛋白的肽， 抑制E2 A蛋白降解。 稳定的组成和 四环素诱导的全长有义平滑肌细胞克隆， 将产生反义UBC-E2 A和UBC-E2 A抑制剂。 他将利用 研究UBC-E2 A对细胞周期限制的影响 点，其调节细胞增殖。 此外，他还将研究 UBC-E2 A对VSMC迁移、粘附和基质蛋白的影响 生产 哈佛医学院心血管生物学实验室 公共卫生部将作为这些研究的中心。 实验室 占地12，000平方英尺，配备了专为 研究细胞和分子生物学。 (End摘要）

DESCRIPTION (Adapted from applicants' abstract) The proliferation and dedifferentiation of vascular smooth muscle cells (VSMC) is a principal component of occlusive vascular disease, which is the leading cause of death in developed countries. As a preventive cardiologist, the candidate's principle focus has been to identify and treat the root causes of atherosclerosis in patients with established cardiovascular disease. As an investigator, he seeks to further the understanding of this disease. The basic helix-loop-helix proteins E12 and E47 promote cellular differentiation and retard cellular proliferation. They are expressed in VSMC. The applicant cloned a novel ubiquitin conjugating enzyme, UBC-E2A, directed against the E12 and E47. The ubiquitin-proteasome system can dramatically alter the cellular phenotype by regulating the degradation of trans-acting factors. By targeting E12 and E47 for turnover, UBC-E2A may promote VSMC proliferation and dedifferentiation. The project herein will characterize E12 and E47 protein degradation in VSMC. First he will map the UBC-E2A binding site on the E2A proteins, then he will determine the role of E2A protein phosphorylation on their degradation. This analysis will permit the design of a peptide to compete with UBC-E2A for binding the E2A proteins and inhibit E2A protein degradation. Stable constitutive and tetracycline-inducible smooth muscle cell clones of full-length sense and antisense UBC-E2A and UBC-E2A inhibitor will be generated. He will use these clones to study the effect of UBC-E2A on the cell cycle restriction point, which regulates cellular proliferation. In addition, he will study the effects of UBC-E2A on VSMC migration, adhesion, and matrix protein production. The Cardiovascular Biology Laboratory of the Harvard School of Public Health will serve as the site for these studies. The laboratory encompasses 12,000 square feet and is equipped with instruments designed for research in cellular and molecular biology. (End of Abstract)