Cell cycle regulation by Ca2+ and its physiological role in yeast

Ca2+对细胞周期的调节及其在酵母中的生理作用

• 批准号: 14206012
• 负责人: MIYAKAWA Tokichi
• 金额: $ 32.78万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14206012/
• 关键词: Saccharomyces cerevisiae; Ca2+ signal transduction; Calcineurin; Cell cycle regulation; S-adenosyl methionine; HOG pathway; Cell cycle checkpoint; Protein kinase C; 細胞周期抑制; S-アデノシルホモシステイン; チェックポイント; Ca^<2+>

To investigate cell cycle regulation by Ca2+ signaling in the yeast Saccharomyces cerevisiae, we isolated many mutants defective in this process and molecular genetic analysis of each mutant was performed. Major outcome are as follows.(1)The isolation of the mutants and their genetic characterization.In addition to the previously characterized 15 complementation groups we further identified 7 new complementation groups to have 22 groups in total.(2)Molecular genetic characterization of scz13 mutant :scz13 mutation was identified as an allele of SAH1 gene that encodes S-adenosylhomocystein (AdoHcy) hydrolase. We have demonstrated that the cellular levels of AdoHcy and its precursor S-adenosylmethionine were highly elevated and cell growth of this mutant was arrested in G1 phase. Due to the elevation of cellular levels of AdoHcy and AdoMet, the expression levels of CLN2 and SWE1 were strongly suppressed. The results indicated the involvement of AdoHcy and AdoMet in cell cycle regulation. … More (3)Molecular genetic characterization of scz13 mutant :scz6 mutation was identified as an allele of a unique PKC1 gene that encodes protein kinase C in yeast. Pkc1 of yeast is known to be involved in the regulation of Mpk1 MAP kinase cascade, which regulates the synthesis of cell wall. We have shown that the defect of scz6 mutant was in the function different from the previously known Pkc1 function. The novel Pkc1 function is required for the expression of Cln2 G1 cyclin which leading to actin polarization polymerization which in turn leads to polar cell growth.(4)Growth regulation that involves calcineurin.We found that calcineurin and high osmolarity glycerol (HOG) pathway antagonize in the regulation of cell growth. More specifically, calcineurin is required for the inhibition of actin polarization at the bud site, and HOG pathway is required for the positive regulation of the formation of the bud after actin has polarized. This mechanism is required for the regulation of bud formation under the conditions of the changes in osmolarity of the environment. Less

为了研究Ca 2+信号在酿酒酵母细胞周期调控中的作用，我们分离了许多在这一过程中有缺陷的突变体，并对每个突变体进行了分子遗传学分析。主要成果如下。（1）突变体的分离及其遗传学特征：除已鉴定的15个互补群外，又鉴定了7个新的互补群，共22个互补群。（2）scz 13突变体的分子遗传学特征：scz 13突变为编码S-腺苷高半胱氨酸水解酶的SAH 1基因的等位基因。我们已经证明，该突变体的细胞中的Hcy及其前体S-腺苷甲硫氨酸的水平高度升高，并且细胞生长被阻止在G1期。由于细胞内同型半胱氨酸和蛋氨酸水平的升高，CLN 2和SWE 1的表达水平被强烈抑制。结果表明，Hcy和Met参与细胞周期调控。 ...更多信息 （3）scz 13突变体的分子遗传学特征：scz 6突变被鉴定为酵母中编码蛋白激酶C的唯一PKC 1基因的等位基因。已知酵母的Pkc 1参与调节Mpk 1 MAP激酶级联反应，其调节细胞壁的合成。我们已经证明scz 6突变体的缺陷在于与先前已知的Pkc 1功能不同的功能。新的Pkc 1功能是Cln 2 G1细胞周期蛋白表达所必需的，这导致肌动蛋白极化聚合，这反过来又导致极性细胞生长。（4）钙调神经磷酸酶（calcineurin）参与的细胞生长调控：我们发现钙调神经磷酸酶（calcineurin）和高渗甘油（high osmolarity glycerol，HOG）通路在细胞生长调控中相互拮抗。更具体地说，钙调磷酸酶是抑制肌动蛋白极化所必需的，而HOG途径是肌动蛋白极化后正调控芽形成所必需的。这种机制是在环境渗透压变化的条件下调节芽形成所必需的。少

项目成果

Koji Kakugawa et al.: "Purification and characterization of a lipase from the glycolipid-producing yeast Kurtzumanomyces sp. I-11"Biosci. Biotechnol. Biochem.. 66. 978-985 (2002)

Koji Kakukawa 等人：“来自糖脂生产酵母 Kurtzumanomyces sp. I-11 的脂肪酶的纯化和表征”Biosci。

Mizunuma et al.: "Involvement of S-adenosylmethionine in G1 cell-cycle regulation in Saccharomyces cerevisiae"Proc.Natl.Acad.Sci.USA. 101(印刷中). (2004)

Mizunuma 等人：“S-腺苷甲硫氨酸参与酿酒酵母 G1 细胞周期调节”Proc.Natl.Acad.Sci.USA 101（印刷中）。

Yuji Miyarnoto et al.: "Identification of Saccharomyces cerevisiae isoeucyl tRNA synthetase as target of G-1-specific inhibitor revreromycin A"J. Biol. Chem.. 277. 28810-28814 (2002)

Yuji Miyarnoto 等人：“鉴定酿酒酵母异亮氨酰 tRNA 合成酶作为 G-1 特异性抑制剂 Revreromycin A 的靶标”J.

Daisuke Morita et al.: "Rpf2p, an evolutionarily conserved protein, interacts with ribosomal protein L11 and is essential for the processing of 27 SB pre-rRNA to 25 SrRNA"J. Biol. Chem.. 277. 28780-28786 (2002)

Daisuke Morita 等人：“Rpf2p 是一种进化上保守的蛋白质，与核糖体蛋白 L11 相互作用，对于 27 SB pre-rRNA 加工成 25 SrRNA 至关重要”。

Koji Kakugawa et al.: "Cloning, characterization, and expression of cDNA encoding a lipase from Kurtzumanomyces sp. I-11"Biosci. Biotechnol. Biochem.. 66. 1328-1336 (2002)

Koji Kakukawa 等人：“编码来自 Kurtzumanomyces sp. I-11 的脂肪酶的 cDNA 的克隆、表征和表达”Biosci。