ANALYSIS OF THE GENE PROMOTERS OF HUMAN BONE MORPHOGENETIC PROTEINS

骨形态发生蛋白（Bone Morphogenetic Proteins，BMPs）是最早发现的诱导骨和软骨异质性的因子，目前已被证实在胚胎发育过程中的器官形成中发挥重要作用。还已知BMP的氨基酸序列从果蝇到哺乳动物是很保守的。迄今为止，BMP根据其氨基酸的同源性被分为几个亚类。然而，鲜为人知的是，每个BMP的生理作用和功能共享。在这项研究中，我们试图通过分析每个BMP基因的启动子区，而不管最近的分类系统的基础上氨基酸同源性，以表征每个BMP的功能共享。对启动子5 ′侧翼区的研究主要是为了研究特定目的基因的表达调控机制。通过启动子区域的包容性分子克隆，我们将启动子研究扩展到“重新”分类一组共享相似氨基酸序列的基因。此外，我们已经开发了一个独特的和敏感的原位杂交系统，使用聚合酶链反应（PCR）产生的单链DNA探针，并使用该技术来研究每个BMP基因是如何调控的转录因子，包括Hox基因。在本研究期间，我们特别对BMP-5和BMP-6基因启动子在正常和前列腺癌病理状态下的特性进行了研究，发现BMP-6基因在表观遗传学上受到Sp1转录因子结合位点周围CpG甲基化的调控。另一方面，BMP-5缺乏此类富含CpG的区域，并且由Hox基因识别的重复ATTA基序调控。作为一项规则，我们进行了我们的研究，使分子生物学测定总是与形态学分析。我们在各种国际和国内会议上展示了我们的数据，并在学术期刊上发表了大量科学论文。

Bone morphogenetic proteins (BMPs), first identified as factors inducing heterogeneous bone and cartilaginous tissues, are now known to play important roles in organogenesis during embryonic development. The amino acids sequences of BMPs are also known to be well conserved from Drosophila to mammalian. So far, BMPs are classified into several subgroups according to their amino acids holomogy. Little is known, however, about the physiologic role and functional sharing of each BMP.In this study, we tried to characterize each BMP's functional sharing by analyzing promoter region of the each BMP gene regardless of the recent classification system based on amino acid homology. The characterization of 5'-flanking region, a promoter structure, is now mostly for the study of the regulation mechanism of gene expression of the specific target gene. By inclusive molecular cloning of the promoter area, we extended the promoter study to 're'-classify a group of genes sharing similar amino acid sequences. Furthermore, we have developed an unique and sensitive in situ hybridization system using a single-stranded DNA probes yielded by polymerase chain reaction (PCR), and used that technique to study how each BMP gene was regulated by transcriptional factors including Hox genes. During this research period, we especially characterized BMP-5 and BMP-6 gene promoters both in normal and pathologic condition as prostate cancer, and found that BMP-6 gene was epigenetically regulated by CpG methylation around the Sp1 transcriptional factor binding sites. On the other hand, BMP-5 lacks such CpG rich area, and regulated by repeated ATTA motifs recognized by Hox genes. As a rule, we performed our studies so that molecular biological assay always went together with morphological analysis. We have presented our data at various international as well as domestic meetings, and publish a lot of scientific papers for academic journals.