Alterations of O-glycosylation, cell wall, and mitochondrial metabolism in Kluyveromyces lactis cells defective in KlPmr1p, the Golgi Ca(2+)-ATPase.
Alterations of O-glycosylation, cell wall, and mitochondrial metabolism in Kluyveromyces lactis cells defective in KlPmr1p, the Golgi Ca(2+)-ATPase.
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DOI：
10.1016/j.bbrc.2004.04.127
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发表时间：
2004-06
影响因子：
3.1
通讯作者：
F. Farina;D. Uccelletti;P. Goffrini;R. A. Butow;C. Abeijon;C. Palleschi
中科院分区：
文献类型：
--
作者：
F. Farina;D. Uccelletti;P. Goffrini;R. A. Butow;C. Abeijon;C. Palleschi
在酵母中，高尔基体的P型钙-ATPase，Pmr1p，是钙稳态中最重要的参与者。在乳酸克鲁维酵母中，KlPMR1失活会导致多效性表型，包括N-糖基化减少和细胞壁形态发生改变。为了研究KlPMR1失活后乳酸克雷伯菌的生理特性，利用包含所有酿酒酵母编码序列的微阵列进行了研究。O-糖基化的改变与K1PMT2的抑制一致，并在O-糖链中确定了末端的N-乙酰氨基葡萄糖。Klpmr1Δ细胞PIRS蛋白表达增强，提示Klpmr1细胞对细胞壁弱化的反应发生在Klpmr1细胞中。我们发现参与乙酰-辅酶A合成的KlPDA1和KlACS2基因过表达，而参与呼吸代谢的KlIDP1、KlACO1和KlSDH2基因表达下调。突变细胞的氧耗量和琥珀酸脱氢酶活性也有增加。所描述的方法突出了KlPMR1在能源生产过程中出人意料的参与。
In yeast the P-type Ca2+-ATPase of the Golgi apparatus, Pmr1p, is the most important player in calcium homeostasis. In Kluyveromyces lactis KlPMR1 inactivation leads to pleiotropic phenotypes, including reduced N-glycosylation and altered cell wall morphogenesis. To study the physiology of K. lactis when KlPMR1 was inactivated microarrays containing all Saccharomyces cerevisiae coding sequences were utilized. Alterations in O-glycosylation, consistent with the repression of KlPMT2, were found and a terminal N-acetylglucosamine in the O-glycans was identified. Klpmr1Δ cells showed increased expression of PIRs, proteins involved in cell wall maintenance, suggesting that responses to cell wall weakening take place in K. lactis. We found over-expression of KlPDA1 and KlACS2 genes involved in the Acetyl-CoA synthesis and down-regulation of KlIDP1, KlACO1, and KlSDH2 genes involved in respiratory metabolism. Increases in oxygen consumption and succinate dehydrogenase activity were also observed in mutant cells. The described approach highlighted the unexpected involvement of KlPMR1 in energy-yielding processes.