Snf1/AMPK promotes SBF and MBF-dependent transcription in budding yeast
Snf1/AMPK promotes SBF and MBF-dependent transcription in budding yeast
复制标题
DOI：
10.1016/j.bbamcr.2013.09.014
复制
发表时间：
2013-12-01
影响因子：
5.1
通讯作者：
Coccetti, Paola
中科院分区：
文献类型：
--
作者：
Busnelli, Sara;Tripodi, Farida;Coccetti, Paola
Snf1 是酵母 AMP 激活激酶同源物，调节涉及适应葡萄糖限制和响应细胞应激的多个基因的表达。我们之前证明Snf1与SBF和MBF复合物的调节亚基Swi6相互作用，并激活CLB5转录。在这里，我们报告说，在 2% 葡萄糖中的 α 因子同步化细胞中，Snfl 催化亚基的丢失会损害 SBF 和 MBF 复合物的结合，以及随后将 FACT 复合物和 RNA 聚合酶 II 募集到 G1 基因启动子上。通过使用 SNF1 的类似物敏感等位基因 SNF1(as)(I132G)，编码一种其催化活性在体内被 2-萘甲基吡唑并嘧啶 1 选择性抑制的蛋白质，我们发现 Snfl 催化活性的抑制会影响 G1 基因的表达，导致通过 a 因子处理或淘洗同步于 G1 期的细胞延迟进入 S 期。此外，Snfl 在 Rpb1（RNA 聚合酶 II 大亚基）的免疫复合物中被检测到，并且在 α 因子释放后 20 分钟存在于 SBF 和 MBF 调节基因的启动子和编码区，表明 Snfl 在 Glregulon 转录激活中具有直接作用。 (C) 2013 Elsevier B.V. 保留所有权利。
Snf1, the yeast AMP-activated kinase homolog, regulates the expression of several genes involved in adaptation to glucose limitation and in response to cellular stresses. We previously demonstrated that Snf1 interacts with Swi6, the regulatory subunit of SBF and MBF complexes, and activates CLB5 transcription. Here we report that, in alpha-factor synchronized cells in 2% glucose, the loss of the Snfl catalytic subunit impairs the binding of SBF and MBF complexes and the subsequent recruitment of the FACT complex and RNA Polymerase II to promoters of G1-genes. By using an analog-sensitive allele of SNF1, SNF1(as)(I132G), encoding a protein whose catalytic activity is selectively inhibited in vivo by 2-naphthylmethyl pyrazolopyrimidine 1, we show that the inhibition of Snfl catalytic activity affects the expression of G1-genes causing a delayed entrance into S phase in cells synchronized in G1 phase by a-factor treatment or by elutriation. Moreover, Snfl is detected in immune complexes of Rpb1, the large subunit of RNA Polymerase II, and is present at both promoters and coding regions of SBF- and MBF-regulated genes 20 min after alpha-factor release, suggesting a direct role for Snfl in the activation of the Glregulon transcription. (C) 2013 Elsevier B.V. All rights reserved.