Laboratory of Cell Cycles of Algae, Institute of Microbiology, ASCR, Třeboň, Czech Republic.
PLoS One. 2011;6(5):e19626. doi: 10.1371/journal.pone.0019626. Epub 2011 May 16.
DNA damage is a threat to genomic integrity in all living organisms. Plants and green algae are particularly susceptible to DNA damage especially that caused by UV light, due to their light dependency for photosynthesis. For survival of a plant, and other eukaryotic cells, it is essential for an organism to continuously check the integrity of its genetic material and, when damaged, to repair it immediately. Cells therefore utilize a DNA damage response pathway that is responsible for sensing, reacting to and repairing damaged DNA. We have studied the effect of 5-fluorodeoxyuridine, zeocin, caffeine and combinations of these on the cell cycle of the green alga Scenedesmus quadricauda. The cells delayed S phase and underwent a permanent G2 phase block if DNA metabolism was affected prior to S phase; the G2 phase block imposed by zeocin was partially abolished by caffeine. No cell cycle block was observed if the treatment with zeocin occurred in G2 phase and the cells divided normally. CDKA and CDKB kinases regulate mitosis in S. quadricauda; their kinase activities were inhibited by Wee1. CDKA, CDKB protein levels were stabilized in the presence of zeocin. In contrast, the protein level of Wee1 was unaffected by DNA perturbing treatments. Wee1 therefore does not appear to be involved in the DNA damage response in S. quadricauda. Our results imply a specific reaction to DNA damage in S. quadricauda, with no cell cycle arrest, after experiencing DNA damage during G2 phase.
DNA 损伤是所有生物基因组完整性的威胁。植物和绿藻特别容易受到 DNA 损伤的影响,尤其是由紫外线引起的损伤,因为它们的光合作用依赖于光线。为了植物和其他真核细胞的生存,生物体必须不断检查其遗传物质的完整性,并在受损时立即修复。因此,细胞利用 DNA 损伤反应途径来负责感知、反应和修复受损的 DNA。我们研究了 5-氟脱氧尿苷、zeocin、咖啡因以及它们的组合对绿藻四尾栅藻细胞周期的影响。如果 DNA 代谢在 S 期之前受到影响,细胞会延迟 S 期并经历永久性 G2 期阻滞;zeocin 引起的 G2 期阻滞被咖啡因部分消除。如果 zeocin 处理发生在 G2 期且细胞正常分裂,则不会观察到细胞周期阻滞。CDKA 和 CDKB 激酶调节 S. quadricauda 中的有丝分裂;它们的激酶活性被 Wee1 抑制。在 zeocin 的存在下,CDKA、CDKB 蛋白水平稳定。相比之下,DNA 扰乱处理对 Wee1 的蛋白水平没有影响。因此,Wee1 似乎不参与 S. quadricauda 的 DNA 损伤反应。我们的结果表明,在经历 G2 期的 DNA 损伤后,S. quadricauda 会产生特定的反应,而不会出现细胞周期阻滞。