TRANSFORMING GENES OF AVIAN SARCOMA VIRUSES

转化禽肉瘤病毒的基因

• 批准号: 3168666
• 负责人: LU-HAI WANG
• 金额: $ 8.16万
• 依托单位: MOUNT SINAI SCHOOL OF MEDICINE OF CUNY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-02-01 至 1992-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3168666
• 关键词: Retroviridae disease; Rous sarcoma virus; antiserum; autoradiography; avian sarcoma virus; chickens; chromosome translocation; complementary DNA; gel electrophoresis; gene expression; genetic manipulation; genetic mapping; immunofluorescence technique; laboratory mouse; laboratory rabbit; laboratory rat; molecular cloning; molecular oncology; mutant; nucleic acid sequence; oncogenes; oncogenic virus; point mutation; protein tyrosine kinase; protooncogene; viral carcinogenesis; virus RNA; virus genetics; virus protein

The objective of this proposal is to understand the nature and functions of the transforming genes of avian sarcoma viruses (ASVs) and their normal cellular counter-part proto-oncogenes. The transforming gene of ASV UR2, v-ros, its corresponding cellular gene, c-ros, and the proto-oncogene c-src will be studied. The functional domains of v-ros will be studied by constructing various site-specific mutants, and ros and src recombinants. Synthesis and subcellular location of the ros protein, its interaction with potential cellular substrates and transforming ability will be compared among parental UR2 and the mutants. The expression of c-ros gene in avian and mammalian tissues at various developmental stages will be examined at RNA and protein levels. The c-ros product will be identified and characterized using ros-specific antisera. In situ immunofluorescence will be performed to identify the specific cells in the tissue expressing the c-ros protein. The cDNA clone of c-ros mRNA will be constructed and sequenced to elucidate the complete c-ros product, which appears to mimic growth factor receptor molecules. To identify the sequence differences between c-ros and v-ros that may be responsible for the different transforming potential of the two genes, c-ros virus and c-ros recombinants will be constructed and compared with respect to their gene products and oncogenicity. Chicken c-src is known to direct the synthesis of a 4 kb mRNA which codes for a 60,000 dalton tyrosine-specific protein kinase. In muscle, while the 4 kb RNA is absent, a 3 kb c-src mRNA lacking most the kinase domain is expressed instead. Both the 4 kb and the 3 kb RNAs will be cloned and sequence to elucidate their nature of synthesis. The potential protein product of the 3 kb RNA will be identified by using antisera against peptides decoded from the nucleotide sequence. The role of this src- related protein in muscle development and function will be studied.

这项建议的目的是了解 禽肉瘤病毒转化基因的功能 (ASV)及其正常的细胞对应原癌基因。 ASV UR2的转化基因v-ros及其相应的 将研究细胞基因c-ros和原癌基因c-src。 V-ros的功能域将通过构建 各种特定位点的突变体，以及ros和src重组体。 RoS蛋白的合成及其亚细胞定位 与潜在细胞底物的相互作用和转化 将比较亲本UR2和突变体之间的能力。 C-ros基因在禽类和哺乳动物组织中的表达 不同的发育阶段将在RNA和 蛋白质水平。C-ros产品将被识别并 用ros特异的抗血清来表征。就地 将进行免疫荧光检查以识别特定的 组织中表达c-ros蛋白的细胞。C DNA克隆 将构建并测序c-ros mRNA，以阐明 完整的c-ros产物，似乎模仿生长因子 受体分子。识别序列差异的方法 在c-ros和v-ros之间可能负责不同的 C-ros病毒和c-ros两个基因的转化潜能 将构建重组体，并将其与 他们的基因产物和致癌性。 已知鸡肉c-src能直接合成4kb的mrna。 它编码一个60,000道尔顿酪氨酸特异的蛋白激酶。 在肌肉中，当4kb的rna缺失时，3kb的c-src mRNA. 相反，缺乏大部分的激活域被表达出来。两人都是4人 Kb和3kb的RNA将被克隆并测序以阐明 它们的合成本质。3的潜在蛋白质产物 利用多肽抗血清鉴定Kb rna 从核苷酸序列中解码出来的。这个资源中心的角色是- 肌肉发育和功能中的相关蛋白质将 学习。