GENE STRUCTURE AND EF-1A IN THE FUNGUS MUCOR

真菌毛霉的基因结构和 EF-1A

• 批准号: 3271990
• 负责人: PAUL S SYPHERD
• 金额: $ 17.75万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-04-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3271990
• 关键词: developmental genetics; enzyme mechanism; fungal genetics; gene expression; genetic manipulation; genetic transcription; genetic translation; genome; lysine; methylation; molecular cloning; nucleic acid hybridization; nucleic acid sequence; protein biosynthesis; radiotracer; ribosomes; sporogenesis; tissue /cell culture; translation factor

The protein synthesis factor EF-1Alpha of Mucor racemosus hyphae contains 8 or 9 methylated lysines per molecule, while the factor from assexual sporangiospores is non-methylated. During the course of spore germination the specific acitivity increases 6 to 9-fold. This increase in specific activity is accompanied by a constant level of mRNA and a level of EF-1Alpha protein that changes little during the germination period. By isolating and sequencing the EF-1Alpha gene, we found tht it is present in 3 copies in the Mucor genome, each in a different genomic context. This proposal is concerned with two aspects of EF-1Alpha: the role of methylation in "activating" the factor, and the genetic organization, including the control of expression of each gene. We will study the effect of methylation on various EF-1Alpha activities in vitro, locate each methylated site within the protein and purify and study the EF-1Alpha methylases. We will further prepare gene-specific hybridization probes to assess the expression of each gene during several morphogenetic changes (e.g., spore germination, yeast-to-hypha conversions, and sporulation). We also propose to study the 5'flanking regions of each gene to assess the mechanisms controlling transcription.

总状毛霉菌丝蛋白合成因子EF-1 α含有8 或9个甲基化赖氨酸，而来自无性繁殖的因子 孢子囊孢子未甲基化。 在孢子萌发过程中 比活性增加6至9倍。 这一增长具体 活性伴随着恒定的mRNA水平和 EF-1 α蛋白在萌发期变化不大。 通过 分离并测序EF-1 α基因，我们发现它存在于 毛霉基因组中有3个拷贝，每个拷贝在不同的基因组环境中。 这 提案涉及EF-1 Alpha的两个方面： 甲基化在“激活”该因子，和遗传组织， 包括控制每个基因的表达。 我们将研究 甲基化对各种EF-1 α活性的影响， 甲基化位点，并纯化和研究EF-1 α 甲基化酶 我们将进一步制备基因特异性杂交探针， 评估在几个形态发生变化期间每个基因的表达 (e.g.,孢子萌发、酵母-菌丝转化和孢子形成）。 我们 我还建议研究每个基因的5 '侧翼区域，以评估 控制转录的机制。