A Schizosaccharomyces pombe Model of Birt-Hogg-Dube Syndrome

Birt-Hogg-Dube 综合征的粟酒裂殖酵母模型

• 批准号: 7015284
• 负责人: Elizabeth P Henske
• 金额: $ 21.38万
• 依托单位: FOX CHASE CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7015284
• 关键词: Schizosaccharomyces pombe; aminoacid transport; biological signal transduction; cell growth regulation; cellular pathology; disease /disorder etiology; enzyme induction /repression; fungal genetics; gene mutation; genetic disorder; genetic models; genetic screening; leucine; mTOR protein; methionine; molecular pathology; skin disorder; syndrome; tumor suppressor genes

DESCRIPTION (provided by applicant): This R21 application is focused on the BHD gene ortholog in the fission yeast Schizosaccharomyces pombe (S. pombe). Birt-Hogg-Dube (BHD) syndrome is an autosomal dominant disorder characterized by skin hamartomas, lung cysts, spontaneous pneumothorax, and renal cell carcinoma. A germline mutation in the BHD gene was also recently found in a large kindred with dominantly inherited primary spontaneous pneumothorax (PSP) and bullous lung lesions. The cellular pathway through which inactivation of BHD leads to disease is completely unknown. However, hamartomas also develop in patients with germline mutations in at least four other tumor suppressor genes (PTEN, TSC1, TSC2 and LKB1). Loss of the protein products of these genes results in activation of the kinase mTOR. The S. pombe BHD ortholog encodes a protein with high sequence conservation to human BHD. In published work, we demonstrated that S. pombe with deletion of the tsc1+ or tsc2+ genes have defects in amino acid uptake. In preliminary studies, we generated a BHD mutant in S. pombe, delta-bhd. We found that delta-bhd S. pombe are resistant to a toxic analog of methionine, suggesting decreased methionine uptake. Delta-hbhd S. pombe also have a defect in the leucine uptake, which is similar to delta-tsc1, delta-tsc2 and delta-tor1 mutants. Based on these data, our central hypothesis is that the Bhd protein signals through Tor1 in S. pombe. We propose the following specific aims. Aim 1: We will determine whether Bhd functions in the same pathway as Tsc1, Tsc2, and Tor1 in S. pombe. Aim 2: We will determine the effect of disease causing conserved mutations in the bhd+ gene on leucine and methionine uptake. Aim 3: We will identify proteins in the Bhd pathway in yeast that can rescue the methionine uptake defect using genetic screens. Our long- term goal is to use this S. pombe model to elucidate the functions of the human BHD protein. This may provide novel insights into cellular pathways involved in the pathogenesis of cystic lung diseases. Lay summary. Yeast models can provide key information about the most important cellular functions of human disease-associated genes. Birt-Hogg-Dube (BHD) syndrome is a rare disease associated with lung cysts, lung collapse, and kidney cancer. Nothing is currently known about how BHD gene mutations lead to human disease. We will use a novel yeast model of BHD to elucidate the function of the BHD protein.

描述（由申请人提供）：该 R21 申请重点关注裂殖酵母裂殖酵母 (S. pombe) 中的 BHD 基因直向同源物。 Birt-Hogg-Dube (BHD) 综合征是一种常染色体显性遗传疾病，其特征为皮肤错构瘤、肺囊肿、自发性气胸和肾细胞癌。最近还在一个具有显性遗传性原发性自发性气胸 (PSP) 和大疱性肺部病变的大型亲属中发现了 BHD 基因的种系突变。 BHD 失活导致疾病的细胞途径尚不清楚。然而，错构瘤也会发生在至少四种其他抑癌基因（PTEN、TSC1、TSC2 和 LKB1）具有种系突变的患者中。这些基因蛋白质产物的丢失会导致激酶 mTOR 的激活。粟酒裂殖酵母 BHD 直向同源物编码的蛋白质与人类 BHD 具有高度序列保守性。在已发表的工作中，我们证明了 tsc1+ 或 tsc2+ 基因缺失的粟酒裂殖酵母在氨基酸摄取方面存在缺陷。在初步研究中，我们在粟酒裂殖酵母中产生了 BHD 突变体 delta-bhd。我们发现 delta-bhd 粟酒裂殖酵母对甲硫氨酸的有毒类似物具有抗性，表明甲硫氨酸的摄取减少。 Delta-hbhd 粟酒裂殖酵母在亮氨酸摄取方面也存在缺陷，这与 delta-tsc1、delta-tsc2 和 delta-tor1 突变体类似。基于这些数据，我们的中心假设是 Bhd 蛋白在粟酒裂殖酵母中通过 Tor1 发出信号。我们提出以下具体目标。目标 1：我们将确定 Bhd 是否与粟酒裂殖酵母中的 Tsc1、Tsc2 和 Tor1 发挥相同的途径。目标 2：我们将确定引起 bhd+ 基因保守突变的疾病对亮氨酸和蛋氨酸摄取的影响。目标 3：我们将通过基因筛选鉴定酵母 Bhd 途径中的蛋白质，这些蛋白质可以挽救蛋氨酸吸收缺陷。我们的长期目标是利用这种粟酒裂殖酵母模型来阐明人类 BHD 蛋白的功能。这可能为涉及囊性肺疾病发病机制的细胞途径提供新的见解。平铺总结。酵母模型可以提供有关人类疾病相关基因最重要的细胞功能的关键信息。 Birt-Hogg-Dube (BHD) 综合征是一种与肺囊肿、肺塌陷和肾癌相关的罕见疾病。目前对于 BHD 基因突变如何导致人类疾病一无所知。我们将使用 BHD 的新型酵母模型来阐明 BHD 蛋白的功能。