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Isolation and characterization of Peutz-Jeghers Syndrome Gene

黑斑息肉综合征基因的分离和表征

基本信息

批准号：
10671180
负责人：
KOYAMA Kumiko
金额：
$ 1.92万
依托单位：
Japanese Foundation for Cancer Research (1999)Osaka University (1998)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671180/
关键词：
PJS STK11 LKB1 Mutation search APCL 53BP2 EB1 EB3 Mutation search Peutz-Jeghers syndrome LKB1遺伝子遺伝子診断 PCR-SSCP法

项目摘要

1. We isolated a novel gene, APCL that showed significant homology to the adenomatous polyposis coli (APC) tumor suppressor gene. This novel gene located on chromosome 19p13.3, which region is STK11/LKB1 gene that cause of Peutz-Jeghers Syndrome (PJS). APCL encodes a protein of 2303 amino acids that is expressed specifically in the brain. The heptad-repeat domain found in the APC protein is well conserved in APCL (45% of the amino acids are identical) ; therefore APCL is likely also to form homo- or hetero- dimers Moreover, since the Armadillo domain is also well conserved (76% identical), both proteins may interact with the same or similar molecular entities. The central portion of APCL consists of five copies of a 20-amino-acid motif and we showed earlier that through this domain APCL could interact with beta-catenin and deplete its intracellular concentration. However, as the C-terminus of APCL protein bears little similarity to that of APC (only 13% identical amino acids), that par … More t of the molecule can be presumed to interact with different proteins ; if so, APCL would possess at least some functions distinct from those of APC. Using a yast two-hybrid system, we attempted to isolate proteins that might associate with the unique COOH-terminus of APCL. Among 166 cDNA clones isolated from a human fetal-brain cDNA library as candidates for interaction with APCL, 32 encoded parts of p53-binding protein 2 (53BP2), a molecule that interacts with p53 and Bcl2, six included an identical sequence with significant homology to EB1, a protein known to bind to APC.2. We investigated the entrie cording of STK11 in 68 patients of 20 unrelated PJS families by the PCR-SSCP method and PCR-direct sequence analysis, and found 15 different, mutations among 14 of those families. One nonsense mutation and eight different frameshift mutations, all of which would cause truncation of the gene product. Moreover three carried intronic mutations at or adjacent to the consensus dinucleotide sequence of splice-acceptor or-donor sites, which were likely to lead to aberrantsplicing. Less

1.我们分离到一个新的基因，APCL，表现出显着的同源性腺瘤性息肉病结肠（APC）肿瘤抑制基因。该新基因定位于染色体19p13.3，该区域为导致Peutz-Jeghers综合征（PJS）的STK 11/LKB 1基因。APCL编码一种2303个氨基酸的蛋白质，在大脑中特异性表达。在APC蛋白中发现的七肽重复结构域在APCL中是很保守的（45%的氨基酸是相同的）;因此APCL也可能形成同源或异源二聚体。此外，由于Armadillo结构域也是很保守的（76%相同），两种蛋白质可以与相同或相似的分子实体相互作用。APCL的中心部分由五个拷贝的20个氨基酸基序组成，我们之前表明，通过这个结构域，APCL可以与β-连环蛋白相互作用，并耗尽其细胞内浓度。然而，由于APCL蛋白的C-末端与APC蛋白的C-末端几乎没有相似性（仅13%相同的氨基酸），因此， ...更多信息可以推测分子的t与不同的蛋白质相互作用;如果是这样的话，APCL将具有至少一些不同于APC的功能。利用酵母双杂交系统，我们试图分离可能与APCL独特的COOH-末端相关的蛋白质。从人胎脑cDNA文库中分离的166个cDNA克隆作为与APCL相互作用的候选者，其中32个编码p53结合蛋白2（53 BP 2）的部分，该分子与p53和Bcl 2相互作用，6个包括与EB 1具有显著同源性的相同序列，EB 1是已知与APC结合的蛋白。应用PCR-SSCP和PCR-direct sequence分析方法，对20个PJS家系的68例患者进行了STK 11基因分型，在14个家系中发现了15种不同的突变。一个无义突变和八个不同的移码突变，所有这些都会导致基因产物的截短。此外，有三个在剪接受体或剪接供体位点的共有二核苷酸序列或其附近携带内含子突变，这可能导致异常剪接。少