MECHANISM OF TRANSFORMATION BY THE V-MYB ONCOGENES

V-MYB 癌基因的转化机制

• 批准号: 2330737
• 负责人: Joseph Steven Lipsick
• 金额: $ 25.74万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 2000-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2330737
• 关键词: DNA binding protein; Schizosaccharomyces pombe; cell cycle proteins; cell growth regulation; chemical binding; chick embryo; complementary DNA; fungal genetics; gene expression; gene mutation; genetic mapping; genetic transcription; laboratory rabbit; molecular biology; mutant; neoplastic transformation; nucleic acid sequence; oligonucleotides; oncogenes; phosphorylation; protein structure function; tissue /cell culture; viral leukemogenesis

The v-myb oncogene causes acute monoblastic leukemia in chickens and induces the leukemic transformation of normal myelomonocytic cells in culture. During the previous funding period, we demonstrated that the ability of the v-Myb protein to activate transcription of a model reporter gene correlated well with its ability to cause leukemic transformation. We also established a number of assays for transcriptional activation and sequence-specific DNA binding by v-Myb and used them to map several functional domains within the v-Myb protein by a combination of biochemical and genetic analyses. c- myb, the normal cellular gene from which v-myb arose, is essential for vertebrate hematopoiesis. Genes related to myb have also been identified in insects, plants, cellular slime molds, and yeasts. Recently, it has been shown that cdc5, a cell division cycle gene of the fission yeast Schizosaccharomyces pombe, encodes a protein with significant similarity to c-Myb. These results suggest that a better understanding of transformation by v-Myb may have broader implications in understanding the basic cell cycle machinery common to all eukaryotes. We have three major goals for the next five years of research on this project. First, we would like to understand in greater molecular detail which features of the v-Myb protein are essential for leukemic transformation. Second, we would like to identify and characterize the other cellular components with which v-Myb interacts. Third, we have developed a model of how the function of the Cdc5 protein in S. pombe relates to tissue-specific transformation by v-Myb and would like to test it experimentally. Therefore, our specific aims for the next funding period are as follows; 1.) To determine which residues of v-Myb are essential for DNA-binding, transcriptional activation, and leukemic transformation. 2.) To identify host cell proteins and DNA sequences which interact directly with v-Myb. 3.) To test the ability of v-Myb to regulate the cell cycle in leukemic cells and in fission yeast.

v-myb癌基因导致鸡的急性单核细胞白血病， 诱导正常骨髓单核细胞的白血病转化， 文化 在上一个资助期间，我们证明了 v-Myb蛋白激活模型报告基因转录的能力 该基因与其导致白血病转化的能力密切相关。 我们 还建立了许多转录激活的测定法， 序列特异性DNA结合的v-Myb，并使用它们来映射几个 v-Myb蛋白质内的功能结构域通过生物化学的组合， 基因分析。 c-myb是产生v-myb的正常细胞基因， 脊椎动物造血 与myb相关的基因也被鉴定出来 在昆虫植物细胞黏菌和酵母中。 近日更 已经证明，裂殖酵母的细胞分裂周期基因cdc 5 粟酒裂殖酵母，编码一种与 c-Myb 这些结果表明，更好地理解转化 通过v-Myb可能有更广泛的意义，在理解基本细胞， 所有真核生物共有的循环机制。 我们在未来五年的研究中有三个主要目标 项目 首先，我们想更详细地了解分子 v-Myb蛋白的哪些特征对于白血病是必需的？ 转型 第二，我们要确定和描述 与v-Myb相互作用的其他细胞组分。 三是 开发了一个模型，研究了Cdc 5蛋白在S. pombe 涉及v-Myb的组织特异性转化，并希望测试 实验性的。 因此，我们下一次筹资的具体目标是 期间如下; 1.）的人。 为了确定v-Myb的哪些残基对于DNA结合是必需的， 转录激活和白血病转化。 2.）的情况。 鉴定宿主细胞蛋白质和DNA序列， 直接注射v-Myb 3.）第三章 检测v-Myb对白血病细胞周期的调节能力， 细胞和裂变酵母中。