Fleischmann Jacob, Rocha Miguel A
Department of Medicine, Greater Los Angeles VA Healthcare System, Los Angeles, California, United States of America; Research Division, Greater Los Angeles VA Healthcare System, Los Angeles, California, United States of America; Department of Medicine, David Geffen School of Medicine at UCLA, Los Angeles, California, United States of America.
Research Division, Greater Los Angeles VA Healthcare System, Los Angeles, California, United States of America.
PLoS One. 2015 May 6;10(5):e0124430. doi: 10.1371/journal.pone.0124430. eCollection 2015.
Candida albicans is an important polymorphic human pathogen. It can switch from a unicellular yeast form to germinating hypha, which may play a role in making it the successful pathogen it is. This hyphal transformation can be triggered by various extracellular stimuli, the most potent one being serum from any source. We have previously reported that Candida albicans transiently polyadenylates portions of both the large and small subunits of ribosomal RNA, shortly after serum exposure. Northern blots at the same time suggested that serum might induce a decrease in total ribosomal RNA. We have carried out a number of experiments to carefully assess this possibility and now report that serum significantly reduces ribosomal RNA in Candida albicans. Fluorometric measurements, Northern blotting and quantitative RT-PCR, have all confirmed this decrease. Timed experiments show that serum induces this decrease rapidly, as it was seen in as early as five minutes. Cell mass is not decreased as total cellular protein content remains the same and metabolic activity does not appear to slow, as assessed by XTT assay, and by the observation that cells form hyphal structures robustly. Another hyphal inducer, N-acetylglucosamine, also caused RNA decrease, but to a lesser extent. We also observed it in non-germinating yeast, such as Candida glabrata. The reason for this decrease is unknown and overall our data suggests that decrease in rRNA does not play a causal role in hyphal transformation. Rapid and significant decrease in a molecule so central to the yeast's biology is of some importance, and further studies, such as its effect on protein metabolism, will be required to better understand its purpose.
白色念珠菌是一种重要的多态性人类病原体。它可以从单细胞酵母形式转变为发芽菌丝,这可能在使其成为成功的病原体方面发挥作用。这种菌丝转变可由各种细胞外刺激引发,其中最有效的是任何来源的血清。我们之前报道过,白色念珠菌在暴露于血清后不久,核糖体RNA的大亚基和小亚基部分会短暂地多聚腺苷酸化。同时进行的Northern印迹分析表明,血清可能会导致总核糖体RNA减少。我们进行了一系列实验来仔细评估这种可能性,现在报告血清会显著降低白色念珠菌中的核糖体RNA。荧光测量、Northern印迹分析和定量RT-PCR均证实了这种减少。定时实验表明,血清能迅速诱导这种减少,早在五分钟时就已观察到。细胞质量并未减少,因为总细胞蛋白含量保持不变,并且通过XTT测定以及观察到细胞能强劲地形成菌丝结构来评估,代谢活性似乎也没有减慢。另一种菌丝诱导剂N-乙酰葡糖胺也会导致RNA减少,但程度较小。我们在非发芽酵母如光滑念珠菌中也观察到了这种情况。这种减少的原因尚不清楚,总体而言我们的数据表明rRNA的减少在菌丝转变中并不起因果作用。对于酵母生物学如此核心的一种分子迅速且显著的减少具有一定重要性,需要进一步研究,例如其对蛋白质代谢的影响,以更好地理解其目的。