SPATIAL PATTERNS OF BUDDING IN YEAST

酵母出芽的空间模式

• 批准号: 2187319
• 负责人: JOHN S CHANT
• 金额: $ 17.34万
• 依托单位: HARVARD 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 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2187319
• 关键词: cell cycle; cell type; cellular polarity; developmental genetics; fungal genetics; gene deletion mutation; gene expression; gene mutation; genetic transcription; goats; growth /development; immunoelectron microscopy; immunofluorescence technique; laboratory rabbit; molecular cloning; nucleic acid sequence; protein biosynthesis; yeasts

Bud formation in yeast affords the opportunity to study simple patterns of spatial organization in a system with well developed genetics, molecular genetics, cell biology and biochemistry. Yeast cells can choose sites for budding in two different spatial patterns. a cells and alpha cells form buds in an axial pattern in which buds are initiated at the mother- daughter junction of the previous division. In the bipolar pattern, a/alpha cells are restricted to bud at their poles; the daughter cell (bud of the previous division) exhibits a strong tendency to bud away from its mother. The long term objective of this project is to understand at the molecular level the machinery responsible for producing the two patterns. Such an understanding requires determining the subcellular location of all proteins involved, knowing the interactions between components, and deciphering the mechanism by which each protein functions, be it a kinase, nucleotide-binding protein or some other type of molecule. Within the scope of this grant proposal, questions which will be addressed are the following: 1) What is the source of axial spatial information? It appears that BUD3 protein is central to furnishing axial information, since it assembles in the mother-bud neck i. e., axial positions. What proteins BUD3 assembles upon, what proteins BUD3 instructs and how BUD3 localization is temporally regulated will be explored. BUD4 , the other axial-specific gene, will be cloned, sequenced, and deleted from the genome. The expression of BUD4 will be examined as the possible basis of cell-type control of budding pattern, and antibodies will be raised to determine its subcellular location. Attempts will be made to identify and analyze additional axial- specific BUD genes. 2) What is the source of bipolar information? An extensive genetic screen will be performed to search for bipolar-specific genes. Interesting genes will be cloned, and analyzed with the goal of this granting period being to localize the products within the cell. 3) What is the basis of cell-type control? BUD4 transcription and the modification state of BUD proteins (especially BUD3) will be examined as the potential basis for cell-type control. The understanding of the mechanisms of budding pattern, that is derived from this work, is of general interest because formation of a bud involves the assembly of cell polarity towards the bud site, and because the plane of cell division directly follows the mother-bud axis. Orientation of an axis of cell polarity and regulation of cell division planes are recurring themes during the development and functioning of multicellular organisms. For example, killing of target cells by T-cells involves polarization of the T-cell towards its target, and specification of cell fates during mouse development is determined by the orientation of cell divisions.

酵母中的芽形成提供了研究简单模式的机会 在具有良好发达遗传学的系统中的空间组织，分子 遗传学，细胞生物学和生物化学。酵母单元可以选择地点 以两种不同的空间模式萌芽。细胞和α细胞形成 在轴向图案的芽中，在母亲处启动芽 上一个部门的女儿交界处。在双极模式中 A/Alpha细胞仅限于杆子的芽；女儿细胞（芽 以前的部门）表现出强烈的趋势 母亲。该项目的长期目标是在 分子水平负责产生这两种模式的机械。 这样的理解需要确定所有的亚细胞位置 涉及的蛋白质，知道组件之间的相互作用以及 破译每种蛋白质功能的机制，无论是激酶， 核苷酸结合蛋白或某些其他类型的分子。在 该赠款提案的范围，将要解决的问题是 下列的： 1）轴向空间信息的来源是什么？看来Bud3 蛋白质是提供轴向信息的核心，因为它在 母亲脖子我。例如，轴向位置。什么蛋白质芽3组装 在上面，蛋白质Bud3指示哪些蛋白质以及Bud3定位在时间上的定位方式 受监管将被探讨。 Bud4，另一个轴向特异性基因将是 克隆，测序和从基因组中删除。芽4的表达 将被检查为细胞类型控制的可能基础 图案和抗体将升高以确定其亚细胞 地点。将尝试识别和分析其他轴向 特定的芽基因。 2）双极信息的来源是什么？广泛的遗传筛查 将进行搜索双极特异性基因。有趣的基因 将被克隆，并以这个授予期的目的进行分析 将产品定位在单元格中。 3）细胞类型控制的基础是什么？ Bud4转录和 芽蛋白的修饰状态（尤其是芽3）将被检查为 细胞类型控制的潜在基础。 对出现模式机制的理解，这是得出的 从这项工作中，人们普遍兴趣，因为芽的形成涉及 细胞极性朝向芽位的组装，因为平面 细胞分裂的直接遵循母体轴。 an的方向 细胞极性轴和细胞分裂平面的调节轴是重复的 多细胞生物的开发和功能期间的主题。 例如，T细胞杀死靶细胞涉及 T细胞朝向其目标，并在细胞命运的规范 小鼠发育取决于细胞分裂的方向。