A Schizosaccharomyces pombe Model of Birt-Hogg-Dube Syndrome

描述（由申请人提供）：该R21申请集中于裂殖酵母粟酒裂殖酵母（Schizosophyomycespombe）（S. pombe）。Birt-Hogg-Dube（BHD）综合征是一种常染色体显性遗传疾病，以皮肤错构瘤、肺囊肿、自发性气胸和肾细胞癌为特征。最近在一个显性遗传的原发性自发性气胸（PSP）和大疱性肺病变的大家族中也发现了BHD基因的种系突变。BHD失活导致疾病的细胞途径完全未知。然而，错构瘤也发生在至少四种其他肿瘤抑制基因（PTEN，TSC 1，TSC 2和LKB 1）发生种系突变的患者中。这些基因的蛋白质产物的丢失导致激酶mTOR的激活。色葡萄粟酒裂殖酵母BHD直向同源物编码与人BHD具有高度序列保守性的蛋白质。在已发表的工作中，我们证明了S。缺失TSC 1+或TSC 2+基因的粟酒酵母具有氨基酸摄取缺陷。在初步研究中，我们在S.蓬贝三角洲公司我们发现δ-bhd S.粟酒裂殖酵母对甲硫氨酸的毒性类似物具有抗性，这表明甲硫氨酸摄取减少。Delta-hbhd S.粟酒裂殖酵母在亮氨酸摄取方面也有缺陷，这与Δ-TSC 1、Δ-TSC 2和Δ-TOR 1突变体相似。基于这些数据，我们的中心假设是Bhd蛋白通过S.粟球。我们提出以下具体目标。目的1：我们将确定Bhd是否与S中的Tsc 1，Tsc 2和Tor 1在相同的通路中起作用。粟球。目的2：我们将确定引起疾病的bhd+基因保守突变对亮氨酸和甲硫氨酸摄取的影响。目的3：我们将确定蛋白质在Bhd途径在酵母中，可以挽救蛋氨酸摄取缺陷使用遗传筛选。我们的长期目标是利用这个S. pombe模型来阐明人BHD蛋白的功能。这可能为囊性肺疾病发病机制中涉及的细胞通路提供新的见解。简单总结。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）综合征是一种罕见的疾病，与肺囊肿，肺萎陷和肾癌有关。目前还不知道BHD基因突变如何导致人类疾病。我们将使用一种新的酵母模型BHD阐明BHD蛋白的功能。

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.