Endocytic Ark/Prk kinases play a critical role in adriamycin resistance in both yeast and mammalian cells.
Endocytic Ark/Prk kinases play a critical role in adriamycin resistance in both yeast and mammalian cells.
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DOI：
10.1158/0008-5472.can-06-3220
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发表时间：
2006-12
期刊：
影响因子：
11.2
通讯作者：
Tsutomu Takahashi;T. Furuchi;A. Naganuma
中科院分区：
文献类型：
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作者：
Tsutomu Takahashi;T. Furuchi;A. Naganuma
为了阐明对阿霉素获得性耐药的机制，我们寻找过表达时使酿酒酵母对阿霉素耐药的基因。我们确定了AKL1，一个功能未知的基因，但被认为是Ark/Prk激酶家族的一员，参与内吞作用的调节，根据其推断的氨基酸序列。在Ark/Prk激酶家族的测试成员（Ark1， Prk1和Akl1）中，Prk1过表达也使酵母细胞对阿霉素产生耐药性。已知Prk1通过磷酸化复合物中的Sla1和Pan1来解离参与内吞作用的Sla1/Pan1/End3复合物。我们发现，与Prk1一样，Akl1促进该复合体中Pan1的磷酸化并降低细胞的内吞能力。Sla1-和End3缺陷酵母细胞也对阿霉素具有耐药性，并且在这些缺陷细胞中过表达Akl1并没有增加阿霉素的耐药程度，这表明Akl1可能通过Sla1/Pan1/End3复合物和Pan1磷酸化的机制，通过降低细胞的内吞能力来降低阿霉素的毒性。我们还发现，过表达人类Ark/Prk家族成员AAK1的HEK293细胞对阿霉素具有耐药性。我们的发现表明，胞吞作用可能参与了阿霉素对酵母和人类细胞的毒性作用机制。
To elucidate the mechanism of acquired resistance to Adriamycin, we searched for genes that, when overexpressed, render Saccharomyces cerevisiae resistant to Adriamycin. We identified AKL1, a gene of which the function is unknown but is considered, nonetheless, to be a member of the Ark/Prk kinase family, which is involved in the regulation of endocytosis, on the basis of its deduced amino acid sequence. Among tested members of the Ark/Prk kinase family (Ark1, Prk1, and Akl1), overexpressed Prk1 also conferred Adriamycin resistance on yeast cells. Prk1 is known to dissociate the Sla1/Pan1/End3 complex, which is involved in endocytosis, by phosphorylating Sla1 and Pan1 in the complex. We showed that Akl1 promotes phosphorylation of Pan1 in this complex and reduces the endocytic ability of the cell, as does Prk1. Sla1- and End3-defective yeast cells were also resistant to Adriamycin and overexpression of Akl1 in these defective cells did not increase the degree of Adriamycin resistance, suggesting that Akl1 might reduce Adriamycin toxicity by reducing the endocytic ability of cells via a mechanism that involves the Sla1/Pan1/End3 complex and the phosphorylation of Pan1. We also found that HEK293 cells that overexpressed AAK1, a member of the human Ark/Prk family, were Adriamycin resistant. Our findings suggest that endocytosis might be involved in the mechanism of Adriamycin toxicity in yeast and human cells.