IDENTIFICATION AND FUNCTIONAL-ANALYSIS OF A DEVELOPMENTALLY-REGULATED EXTRACELLULAR SIGNAL-REGULATED KINASE GENE IN DICTYOSTELIUM-DISCOIDEUM
IDENTIFICATION AND FUNCTIONAL-ANALYSIS OF A DEVELOPMENTALLY-REGULATED EXTRACELLULAR SIGNAL-REGULATED KINASE GENE IN DICTYOSTELIUM-DISCOIDEUM
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DOI：
10.1128/mcb.14.10.6996
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发表时间：
1994-10-01
影响因子：
5.3
通讯作者：
FIRTEL, RA
中科院分区：
文献类型：
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作者：
GASKINS, C;MAEDA, M;FIRTEL, RA
我们从盘基网柄菌中克隆了一种发育调节的丝裂原激活蛋白激酶（细胞外信号调节激酶），命名为 ERK1。使用抗 pTyr 抗体，我们发现 ERK1 在体内酪氨酸上被磷酸化，并且它会磷酸化髓磷脂碱性蛋白。该基因表达两种转录本，一种在营养生长和早期发育期间优先表达，另一种在多细胞阶段诱导表达。使用抗 ERK1 抗体进行的发育蛋白质印迹（免疫印迹）表明 ERK1 存在于整个发育过程中。 ERK1/lacZ 报告基因构建体表明，在多细胞阶段，该基因优先在分散在整个生物体中的细胞亚群中表达，类似于前部样细胞标记物所见的模式。来自可抑制启动子的反义诱变表明 ERK1 对于营养生长至关重要。肌动蛋白 15 启动子或 ERK1 启动子过度表达 ERK1 会导致从蛞蝓阶段开始的形态发生异常。 ERK1 在磷酸酪氨酸磷酸酶 PTP2 无效突变体中的过度表达会导致大量聚集流的产生以及随后的异常形态发生，这表明 ERK1 和 PTP2 之间存在遗传相互作用。这些细胞产生非常大的聚集流，这些聚集流分解成非常小的土堆，并经历异常的形态发生。 ERK1 和 PTP2 之间的遗传相互作用似乎是特异性的，因为 ptp(1-) 无效突变体中 ERK1 的过度表达不会产生相同的表型。我们的结果表明 ERK1 在 D. discoideum 的生长和分化过程中发挥着重要作用。
We have cloned a developmentally regulated mitogen-activated protein kinase (extracellular signal-regulated kinase) from Dictyostelium discoideum designated ERK1. Using anti-pTyr antibodies, we show that ERK1 is phosphorylated on tyrosine in vivo and that it will phosphorylate myelin basic protein. The gene expresses two transcripts, one that is preferentially expressed during vegetative growth and early development and one that is induced during the multicellular stages. Developmental Western blots (immunoblots) using anti-ERK1 antibodies indicate that ERK1 is present throughout development. ERK1/lacZ reporter constructs suggest that, in the multicellular stages, the gene is preferentially expressed in a subpopulation of cells scattered throughout the organism, similar to the pattern seen with anterior-like cell markers. Antisense mutagenesis from a derepressible promoter indicates that ERK1 is essential for vegetative growth. Overexpression of ERK1 from either the Actin 15 promoter or the ERK1 promoter results in abnormal morphogenesis starting at the slug stage. Overexpression of ERK1 in null mutants of the phosphotyrosine phosphatase PTP2 results in the production of large aggregation streams and subsequent abnormal morphogenesis that indicate a genetic interaction between ERK1 and PTP2. These cells produce very large aggregation streams that break up into very small mounds that undergo abnormal morphogenesis. The genetic interaction between ERK1 and PTP2 appears to be specific since overexpression of ERK1 in a ptp(1-) null mutant does not produce the same phenotype. Our results indicate that ERK1 plays an essential role during the growth and differentiation of D. discoideum.