Cullen P J, Sprague G F
Institute of Molecular Biology, University of Oregon, Eugene, OR 97403-1229, USA.
Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13619-24. doi: 10.1073/pnas.240345197.
Haploid yeast invades solid agar in response to nutrient limitation. To decipher the cues that underlie invasion, we have developed a single cell invasive growth assay. Using this assay, as well as the traditional plate-washing assay, we show that invasive growth occurs in response to glucose depletion. In the absence of glucose (or other fermentable sugar), individual cells adopted a nonaxial budding pattern and elongated morphology within the first cell divisions, and invasion into the agar was observed in microcolonies containing as few as 10 cells. In support of this observation, we found that glucose suppressed the hyperinvasive growth morphology of STE11-4, pbs2, hsl7, and RAS2V19 mutations. In addition, removal of glucose from YPD medium caused constitutive invasion in wild-type cells. We tested glucose control proteins for a role in invasion and found that Snf1, a protein required for derepression of glucose-repressed genes, was required for invasive growth. The transcription factor Sip4, which interacts with Snf1 and is induced during the diauxic shift, had an inhibitory role on invasive growth, suggesting that multiple mechanisms are required for glucose depletion-dependent invasion.
单倍体酵母在营养限制时会侵入固体琼脂。为了解析侵袭背后的线索,我们开发了一种单细胞侵袭生长测定法。使用这种测定法以及传统的平板洗涤测定法,我们发现侵袭生长是对葡萄糖耗尽的反应。在没有葡萄糖(或其他可发酵糖)的情况下,单个细胞在最初的细胞分裂中采用非轴向出芽模式并呈现细长形态,并且在含有低至10个细胞的微菌落中观察到侵入琼脂的现象。为支持这一观察结果,我们发现葡萄糖抑制了STE11 - 4、pbs2、hsl7和RAS2V19突变的超侵袭生长形态。此外,从YPD培养基中去除葡萄糖会导致野生型细胞的组成型侵袭。我们测试了葡萄糖调控蛋白在侵袭中的作用,发现Snf1,一种解除葡萄糖抑制基因抑制所必需的蛋白质,是侵袭生长所必需的。转录因子Sip4,它与Snf1相互作用并在双相转换期间被诱导,对侵袭生长具有抑制作用,这表明葡萄糖耗尽依赖性侵袭需要多种机制。