The CATS (FAM64A) protein is a substrate of the Kinase Interacting Stathmin (KIS)
The CATS (FAM64A) protein is a substrate of the Kinase Interacting Stathmin (KIS)
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DOI：
10.1016/j.bbamcr.2013.02.004
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发表时间：
2013-05-01
影响因子：
5.1
通讯作者：
Olalla Saad, Sara Teresinha
中科院分区：
文献类型：
--
作者：
Archangelo, Leticia Froehlich;Greif, Philipp A.;Olalla Saad, Sara Teresinha
CATS蛋白（也称为FAM 64 A和RCS 1）首先被鉴定为一种新的CALM（PICALM）相互作用物，其影响致白血病融合蛋白CALM/AF 10的亚细胞定位。CATS以细胞周期依赖性方式在癌细胞系中高度表达，并由有丝分裂原诱导。CATS被认为是增殖的标志物，已知在细胞分裂期间控制中期到后期的转变。在酵母双杂交筛选中使用CATS作为诱饵，我们鉴定了激酶相互作用Stathmin（MS或UHMK 1）蛋白作为CATS相互作用伴侣。通过GST pull-down、免疫共沉淀和共定位实验证实了CATS和KIS之间的相互作用。用激酶分析法，我们发现CATS是KIS的底物，并将磷酸化位点定位到CATS丝氨酸131（S131）。蛋白质表达分析表明，KIS水平的变化在细胞周期依赖性的方式和在相反的方向，CATS水平。在报告基因测定中，KIS能够增强CATS的转录抑制活性，而不依赖于CATS在S131的磷酸化。CATS和KIS均能拮抗CALM/AF 10的反式激活能力。总之，我们的研究结果表明，CATS与KIS相互作用，并且是KIS的底物，这表明KIS调节CATS的功能。(c)2013 Elsevier B. V.保留所有权利。
The CATS protein (also known as FAM64A and RCS1) was first identified as a novel CALM (PICALM) interactor that influences the subcellular localization of the leukemogenic fusion protein CALM/AF10. CATS is highly expressed in cancer cell lines in a cell cycle dependent manner and is induced by mitogens. CATS is considered a marker for proliferation, known to control the metaphase-to-anaphase transition during the cell division. Using CATS as a bait in a yeast two-hybrid screen we identified the Kinase Interacting Stathmin (MS or UHMK1) protein as a CATS interacting partner. The interaction between CATS and KIS was confirmed by GST pull-down, co-immunopreciptation and co-localization experiments. Using kinase assay we showed that CATS is a substrate of KIS and mapped the phosphorylation site to CATS serine 131 (S131). Protein expression analysis revealed that KIS levels changed in a cell cycle-dependent manner and in the opposite direction to CATS levels. In a reporter gene assay KIS was able to enhance the transcriptional repressor activity of CATS, independent of CATS phophorylation at S131. Moreover, we showed that CATS and KIS antagonize the transactivation capacity of CALM/AF10.In summary, our results show that CATS interacts with and is a substrate for KIS, suggesting that KIS regulates CATS function. (c) 2013 Elsevier B.V. All rights reserved.