Cloning and Initial Functional Characterization of Mlk4α and Mlk4β.

Genomics insights Pub Date : 2011-03-22 eCollection Date: 2011-01-01 DOI:10.4137/GEI.S6092

Vladimir I Kashuba, Elvira V Grigorieva, Sergei M Kvasha, Tatiana V Pavlova, Vladimir Grigoriev, Alexei Protopopov, Olga Kharchenko, Rinat Gizatullin, Alla V Rynditch, Eugene R Zabarovsky

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引用次数: 10

Abstract

We have cloned a novel human mixed-lineage kinase gene, MLK4. Two alternatively spliced forms, MLK4α (580 aa) and MLK4β (1036 aa), have been identified and mapped to chromosomal band 1q42. MLK4 shows high amino acid homology to the kinase catalytic domain of MLK3 (72%), MLK1 (71%) and MLK2 (69%). Strong expression of MLK4 was detected in the human pancreas and kidneys. pCMV-MLK4β c-myc-tagged protein (human) was expressed in the cytoplasm and nucleus of transiently transfected COS-1 cells, while pCMV-MLK4α c-myc-tagged protein (human) was expressed in cytoplasm only. Both MLK4 isoforms reduced the colony formation ability of MCF7 cells by 85%-95% and almost totally suppressed cell proliferation in the CyQUANT cell proliferation assay. Human pCMV-MLK4β transgenic mice expressed the MLK4β in all tissues examined but no phenotypic abnormalities were observed. Thus, in this work, we present the cloning and sequencing of MLK4α and MLK4β for the first time; the data obtained suggest that MLK4 may function as a MAP kinase.

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Mlk4α和Mlk4β基因的克隆及初步功能鉴定

我们克隆了一种新的人类混合谱系激酶基因MLK4。MLK4α (580 aa)和MLK4β (1036 aa)两种选择性剪接形式已被鉴定并定位到染色体带1q42上。MLK4与MLK3(72%)、MLK1(71%)和MLK2(69%)的激酶催化结构域具有高度的氨基酸同源性。在人胰腺和肾脏中检测到MLK4的强表达。pCMV-MLK4β c-myc标记蛋白(人)在瞬时转染的COS-1细胞的细胞质和细胞核中表达，而pCMV-MLK4α c-myc标记蛋白(人)仅在细胞质中表达。在CyQUANT细胞增殖实验中，两种MLK4亚型均使MCF7细胞的集落形成能力降低了85%-95%，并且几乎完全抑制了细胞增殖。人pCMV-MLK4β转基因小鼠在所有组织中均表达MLK4β，但未观察到表型异常。因此，本研究首次对MLK4α和MLK4β进行了克隆和测序;所获得的数据表明MLK4可能具有MAP激酶的功能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Genomics insights

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