Saccharomyces cerevisiae protein phosphatase Ppz1 and protein kinases Sat4 and Hal5 are involved in the control of subcellular localization of Gln3 by likely regulating its phosphorylation state
Saccharomyces cerevisiae protein phosphatase Ppz1 and protein kinases Sat4 and Hal5 are involved in the control of subcellular localization of Gln3 by likely regulating its phosphorylation state
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DOI：
10.1016/j.jbiosc.2010.11.013
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发表时间：
2011-03-01
影响因子：
2.8
通讯作者：
Harashima, Satoshi
中科院分区：
文献类型：
--
作者：
Hirasaki, Masataka;Horiguchi, Masaya;Harashima, Satoshi
缺乏PPZ 1的酿酒酵母突变体编码丝氨酸/苏氨酸蛋白磷酸酶（PPase），对咖啡因敏感。为了阐明Ppz 1在咖啡因抗性中的作用，我们试图系统地鉴定蛋白激酶（PKase），其破坏导致Delta ppz 1破坏物的咖啡因敏感表型的抑制，因为PPZ 1的破坏可能通过增加其磷酸化底物而引起咖啡因敏感性，并且我们推测与Ppz 1共享底物的PKase基因的破坏可以通过绕过必需性来恢复抗性用于底物的去磷酸化。在102个可行的pkase破坏中，SAT 4或HAL 5的破坏抑制了咖啡因敏感性表型，并增加了ENA 1的表达，ENA 1编码Delta ppz 1破坏物的P型ATP酶。因为发现Delta ppz 1破坏物中ENA 1表达的增加被GLN 3的破坏所抑制，所以将Delta ppz 1破坏物中Gln 3的定位和磷酸化与Delta ppz 1 Delta sat 4和Delta ppz 1 Delta hal 5双破坏物中Gln 3的定位和磷酸化进行比较。Gln 3被发现积累在细胞核中的三角洲ppz 1破坏剂，并取消了中断的SAT 4或HAL 5的这种核定位。有趣的是，在Delta ppz 1 Delta sat 4和Delta ppz 1 Delta hal 5破坏物中Gln 3磷酸化水平相对于野生型降低，不依赖于咖啡因。从这些观察，我们得出结论，Ppz 1控制Gln 3的本地化，通过调节其磷酸化状态与Sat 4和Hal 5相结合。(C)2010年，生物技术学会，日本。All rights reserved.
A Saccharomyces cerevisiae mutant lacking PPZ1, encoding a serine/threonine protein phosphatase (PPase), is caffeine-sensitive. To clarify the function of Ppz1 in resistance to caffeine, we attempted systematically to identify protein kinase (PKase) whose disruption lead to suppression of caffeine sensitive phenotype of the Delta ppz1 disruptant since disruption of PPZ1 might cause caffeine sensitivity by increasing its phosphorylated substrates and we presumed that disruption of genes for PKase sharing the substrate with Ppz1 could restore the resistance through bypassing necessity for dephosphorylation of substrates. Among the 102 viable pkase disruptions, disruption of either SAT4 or HAL5 suppressed the caffeine sensitivity phenotype and increased expression of ENA1, encoding a P-type ATPase of the Delta ppz1 disruptant. Because increased expression of ENA1 in the Delta ppz1 disruptant was found to be suppressed by disruption of GLN3, localization and phosphorylation of Gln3 in the Delta ppz1 disruptant was compared to that in the Delta ppz1 Delta sat4 and Delta ppz1 Delta hal5 double disruptants. Gln3 was found to accumulate in the nucleus in the Delta ppz1 disruptant, and this nuclear localization was abolished by disruption of either SAT4 or HAL5. Interestingly, the level of Gln3 phosphorylation in the Delta ppz1 Delta sat4 and Delta ppz1 Delta hal5 disruptants decreased relative to wild type independent of caffeine. From these observations, we conclude that Ppz1 controls Gln3 localization by regulating its phosphorylation state in combination with Sat4 and Hal5. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.