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Pak1 Protein Kinase Regulates Activation and Nuclear Localization of Snf1-Gal83 Protein Kinase

机译：Pak1蛋白激酶调节Snf1-Gal83蛋白激酶的激活和核定位。

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摘要

Three kinases, Pak1, Tos3, and Elm1, activate Snf1 protein kinase in Saccharomyces cerevisiae. This cascade is conserved in mammals, where LKB1 activates AMP-activated protein kinase. We address the specificity of the activating kinases for the three forms of Snf1 protein kinase containing the β-subunit isoforms Gal83, Sip1, and Sip2. Pak1 is the most important kinase for activating Snf1-Gal83 in response to glucose limitation, but Elm1 also has a significant role; moreover, both Pak1 and Elm1 affect Snf1-Sip2. These findings exclude the possibility of a one-to-one correspondence between the activating kinases and the Snf1 complexes. We further identify a second, unexpected role for Pak1 in regulating Snf1-Gal83: the catalytic activity of Pak1 is required for the nuclear enrichment of Snf1-Gal83 in response to carbon stress. The nuclear enrichment of Snf1 fused to green fluorescent protein (GFP) depends on both Gal83 and Pak1 and is abolished by a mutation of the activation loop threonine; in contrast, the nuclear enrichment of Gal83-GFP occurs in a snf1Δ mutant and depends on Pak1 only when Snf1 is present. Snf1-Gal83 is the only form of the kinase that localizes to the nucleus. These findings, that Pak1 both activates Snf1-Gal83 and controls its nuclear localization, implicate Pak1 in regulating nuclear Snf1 protein kinase activity.
机译：Pak1，Tos3和Elm1这三种激酶可激活酿酒酵母中的Snf1蛋白激酶。该级联在哺乳动物中得以保留，其中LKB1激活AMP激活的蛋白激酶。我们针对三种形式的包含β亚基异构体Gal83，Sip1和Sip2的Snf1蛋白激酶激活激酶的特异性。 Pak1是响应葡萄糖限制激活Snf1-Gal83的最重要的激酶，但是Elm1也起着重要的作用。此外，Pak1和Elm1都会影响Snf1-Sip2。这些发现排除了活化激酶和Snf1复合物之间一一对应的可能性。我们进一步确定了Pak1在调节Snf1-Gal83中的第二个出乎意料的作用：Pak1的催化活性是对Snf1-Gal83的碳富集以响应碳胁迫所必需的。与绿色荧光蛋白（GFP）融合的Snf1的核富集依赖于Gal83和Pak1，并被激活环苏氨酸的突变所废除。相反，Gal83-GFP的核富集发生在 snf1 Δ突变体中，并且仅在存在Snf1时才依赖于Pak1。 Snf1-Gal83是定位于细胞核的唯一激酶形式。这些发现Pak1既激活Snf1-Gal83并控制其核定位，也暗示Pak1调节核Snf1蛋白激酶活性。

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