CoPK32 is a novel stress-responsive protein kinase in the mushroom Coprinopsis cinerea
CoPK32 is a novel stress-responsive protein kinase in the mushroom Coprinopsis cinerea
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CoPK32 是蘑菇灰鬼伞 (Coprinopsis cinerea) 中的一种新型应激响应蛋白激酶
DOI：
10.1016/j.bbagen.2011.03.018
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Keisuke Kaneko
中科院分区：
文献类型：
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作者：
遠藤圭太;荒川圭太;藤川清三;遠藤圭太;遠藤圭太;遠藤圭太;K.Endoh;K.Endoh;Keisuke Kaneko
在之前的研究中，我们利用Multi-PK抗体对从Coprinopsis cinerea菌丝体中制备的cDNA文库进行了表达克隆筛选，并检测到多种丝氨酸/苏氨酸蛋白激酶。其中一个分离的克隆CMZ032被发现编码一种假定的丝氨酸/苏氨酸蛋白激酶，称为CoPK32。在本研究中，我们研究了CoPK32的生化特性和生理意义。方法在大肠杆菌中表达scopk32，并检测其生化特性。高渗透胁迫对c生长的影响。并对其菌丝体内源CoPK32活性进行了检测。结果scopk32具有自磷酸化活性，能有效磷酸化外源蛋白底物。CoPK32S是一个比CoPK32短18个氨基酸的剪接变体，其蛋白激酶活性明显低于CoPK32。CoPK32缺失突变体的催化特性表明，CoPK32的c端区域对激酶活性和底物识别很重要。CoPK32在菌丝集落的活跃生长区域高表达。当高渗透胁迫刺激菌丝时，内源CoPK32被显著激活，菌丝生长受到严重抑制。作为众所周知的p38丝裂原活化蛋白激酶抑制剂，SB202190或SB239063消除了高渗透胁迫对CoPK32活性的激活作用。结论scopk32参与了c菌丝体的应激反应途径。对环境压力的快速反应。一般significanceInC。此外，CoPK32等蛋白激酶在参与应激反应的信号转导途径中发挥重要作用。
BackgroundIn a previous study, we conducted an expression cloning screen of a cDNA library prepared from Coprinopsis cinerea mycelia using Multi-PK antibodies and detected a wide variety of Ser/Thr protein kinases. One of the isolated clones, CMZ032, was found to encode a putative Ser/Thr protein kinase designated CoPK32. In the present study, we investigated the biochemical properties and physiological significance of CoPK32.MethodsCoPK32 was expressed inEscherichia coli, and its biochemical properties were examined. The effects of high osmotic stresses on the growth ofC. cinereaand on the endogenous CoPK32 activity in mycelia were also examined.ResultsCoPK32 showed autophosphorylation activity and effectively phosphorylated exogenous protein substrates. CoPK32S, a splice variant that was 18 amino acids shorter than CoPK32, showed much lower protein kinase activity than CoPK32. The catalytic properties of CoPK32 deletion mutants suggested that the C-terminal region of CoPK32 was important for the kinase activity and recognition of substrates. CoPK32 was highly expressed in the actively growing region of the mycelial colony. When mycelia were stimulated by high osmotic stresses, endogenous CoPK32 was markedly activated and the mycelial growth was severely inhibited. The activation of CoPK32 activity by high osmotic stresses was abrogated by SB202190 or SB239063 as well-known inhibitors of p38 mitogen-activated protein kinase.ConclusionsCoPK32 is involved in the stress response pathway in mycelia ofC. cinereain response to environmental stresses.General significanceInC. cinerea, protein kinases such as CoPK32 play important roles in signal transduction pathways involved in stress responses.