Molecular analysis of cell cycle regulation by Ca^<2+>

Ca^<2>细胞周期调节的分子分析

• 批准号: 10460044
• 负责人: MIYAKAWA Tokichi
• 金额: $ 8.13万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10460044/
• 关键词: Ca^<2+> signaling; Saccharomyces cerevisiae; Calcineurin; Cell cycle; Ca^<2+>シグナル; カルシニューリンニューリン; MAPキナーゼ; カルシウム; Ca^<2+>; チェックポイント

We investigated molecular mechanism of G2/M cell cycle regulation by Ca^<2+> signal in Saccharomyces cerevisiae. We isolated mutants with a defect in the Ca^<2+> signaling pathway linked to cell cycle regulation. The mutants were classified to 14 complementation groups.(1) csz7 mutant was analyzed in detail. The scz7 mutation was identified as an allele of MCK1 that encodes a GSK-3 family protein kinase highly conserved among eukaryotes. It was found that Mck1 was involved in the Ca^<2+>-induced destabilization of Hsl1, Negative regulator of Swe1 through the ubiquitin-proteasome pathway. We further revealed the detailed mechanism by which Hsl1 abundance is regulated.(2) scz6 mutant was analyzed in detail. The scz6 mutation was identified as an allele of PKC1 that encodes a unique protein kinase C in the yeast. Interestingly, the scz6 mutation suppressed Ca^<2+> sensitive growth and Ca^<2+>-induced polarized bud growth, but not G2 cell cycle delay. We found a novel pathway of Pkc1, in which Pkc1 plays an important role in the regulation of the transcription of SWE1 And the G1 cyclin CLN1, in addition to the previously characterized pathway that activates the Mpk1 MAP kinase pathway. The transcription factor Swi4 was implicated in the transcriptional regulation of CLN1 and SWE1.scz5 mutant was analyzed in detail. The scz5 mutation was identified as an allele of PDR5 that encodes an ABC transporter implicated in multidrug resistance in the yeast. Pdr5 was suggested to be responsible for Ca^<2+> uptake.

我们研究了Ca^<2+>信号对酿酒酵母G2/M期细胞周期调控的分子机制。我们分离出了与细胞周期调控相关的Ca^2+信号通路缺陷的突变体。将突变体分为14个互补群。(1)对CSZ 7突变体进行了详细分析。scz 7突变被鉴定为MCK 1的等位基因，其编码在真核生物中高度保守的GSK-3家族蛋白激酶。Mck 1通过泛素-蛋白酶体途径参与Ca^<2+>诱导的Swe 1负调节因子Hsl 1的失稳。我们进一步揭示了Hsl 1丰度调节的详细机制。(2)对scz 6突变体进行了详细的分析。scz 6突变被鉴定为PKC 1的等位基因，其编码酵母中独特的蛋白激酶C。有趣的是，scz 6突变抑制了Ca^<2+>敏感性生长和Ca^<2+>诱导的极化芽生长，但不抑制G2期细胞周期延迟。我们发现了一条新的Pkc 1通路，其中Pkc 1在SWE 1和G1期细胞周期蛋白CLN 1的转录调控中起重要作用，除了先前表征的激活Mpk 1 MAP激酶通路的通路之外。研究表明，转录因子Swi 4参与了CLN 1和SWE 1的转录调控。scz 5突变被鉴定为PDR 5的等位基因，其编码与酵母中的多药耐药性有关的ABC转运蛋白。推测Pdr 5可能是Ca^2+摄取的主要途径。

项目成果

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I.C.Farcasanu：“组氨酸渗透酶 (Hip1p) 参与酿酒酵母的锰转运。”

Masaki Mizunuma et al.: "GSK-3 kinase Mck1 and calcineurin coordinately mediates Hsl1 down-regulation by Ca^<2+> in budding yeast"EMBO J.. 20. 1074-1085 (2001)

Masaki Mizunuma 等人：“GSK-3 激酶 Mck1 和钙调神经磷酸酶在芽殖酵母中协调介导 Ca^2 下调 Hsl1”EMBO J.. 20. 1074-1085 (2001)

Rota Tsujii et al.: "Ebp2, yeast homologue of a human protein that interacts with Epstein-Barr virus nuclear antigen 1, is required for pre-rRNA processing and ribosomal subunit assembly"Genes to Cell. 5. 543-553 (2000)

Rota Tsujii 等人：“Ebp2 是一种与 Epstein-Barr 病毒核抗原 1 相互作用的人类蛋白质的酵母同源物，是 rRNA 前体加工和核糖体亚基组装所必需的”Genes to Cell。

Z.Cui: "Functional analysis of the yeast SNQ2 gene encoding a yeast multidrug resistance tranaporter that confers resistance to 4 NQO"Biosci.Biotechnol.Biochem.. 63(1). 162-167 (1999)

Z.Cui：“编码酵母多药抗性转运蛋白的酵母 SNQ2 基因的功能分析，该基因赋予对 4 NQO 的抗性”Biosci.Biotechnol.Biochem.. 63(1)。

Tomoki Hosotani et al.: "PKC1,a protein kinase C homologue of Saccharomyces cerevisiase, participates in microtubule function through the yeast EB1 homologue BIM1"Gene Cells. 6. 775-788 (2001)

Tomoki Hosotani 等人：“PKC1 是酿酒酵母的蛋白激酶 C 同源物，通过酵母 EB1 同源物 BIM1 参与微管功能”基因细胞。