Laboratory of Cancer and Developmental Cell Biology, Van Andel Research Institute, Grand Rapids, Michigan, United States of America.
PLoS One. 2011 Feb 18;6(2):e17165. doi: 10.1371/journal.pone.0017165.
Mitogen-activated protein kinase kinases (MKK or MEK) 1 and 2 are usually treated as redundant kinases. However, in assessing their relative contribution towards ERK-mediated biologic response investigators have relied on tests of necessity, not sufficiency. In response we developed a novel experimental model using lethal toxin (LeTx), an anthrax toxin-derived pan-MKK protease, and genetically engineered protease resistant MKK mutants (MKKcr) to test the sufficiency of MEK signaling in melanoma SK-MEL-28 cells. Surprisingly, ERK activity persisted in LeTx-treated cells expressing MEK2cr but not MEK1cr. Microarray analysis revealed non-overlapping downstream transcriptional targets of MEK1 and MEK2, and indicated a substantial rescue effect of MEK2cr on proliferation pathways. Furthermore, LeTx efficiently inhibited the cell proliferation and anchorage-independent growth of SK-MEL-28 cells expressing MKK1cr but not MEK2cr. These results indicate in SK-MEL-28 cells MEK1 and MEK2 signaling pathways are not redundant and interchangeable for cell proliferation. We conclude that in the absence of other MKK, MEK2 is sufficient for SK-MEL-28 cell proliferation. MEK1 conditionally compensates for loss of MEK2 only in the presence of other MKK.
丝裂原活化蛋白激酶激酶(MKK 或 MEK)1 和 2 通常被视为冗余激酶。然而,在评估它们对 ERK 介导的生物学反应的相对贡献时,研究人员依赖于必要性而非充分性的测试。作为回应,我们使用致死毒素(LeTx)、一种炭疽毒素衍生的泛 MKK 蛋白酶和基因工程蛋白酶抗性 MKK 突变体(MKKcr)开发了一种新的实验模型,以测试 MEK 信号在黑色素瘤 SK-MEL-28 细胞中的充分性。令人惊讶的是,ERK 活性在表达 MEK2cr 的 LeTx 处理细胞中持续存在,但在表达 MEK1cr 的细胞中不存在。微阵列分析揭示了 MEK1 和 MEK2 的非重叠下游转录靶标,并表明 MEK2cr 对增殖途径有实质性的挽救作用。此外,LeTx 可有效抑制表达 MKK1cr 但不表达 MEK2cr 的 SK-MEL-28 细胞的增殖和非锚定依赖性生长。这些结果表明,在 SK-MEL-28 细胞中,MEK1 和 MEK2 信号通路不是冗余的,不能相互替代以促进细胞增殖。我们得出的结论是,在没有其他 MKK 的情况下,MEK2 足以促进 SK-MEL-28 细胞的增殖。只有在存在其他 MKK 的情况下,MEK1 才会有条件地补偿 MEK2 的缺失。