Tirumani S, Gothandam K M, J Rao Basuthkar
B-202, Department of Biological Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Colaba, Mumbai, 400005, India.
School of Bio Sciences and Technology, VIT University, Vellore, Tamil Nadu, 632014, India.
Protoplasma. 2019 Jan;256(1):117-130. doi: 10.1007/s00709-018-1283-4. Epub 2018 Jul 10.
Carbon concentrating mechanism (CCM) and photorespiration (PR) are interlinked and co-regulated in Chlamydomonas reinhardtii, but conditions where co-regulation alters are not sufficiently explored. Here, we uncover that PR gene transcripts, like CCM transcripts, are induced even in the dark when both processes are not active. Such diurnal cycles show that transcript levels peak in the middle of 12 h day, decline by early part of 12-h dark followed by their onset again at mid-dark. Interestingly, the onset in the mid-dark phase is sensitive to high CO, implying that the active carbon sensing mechanism operates even in the dark. The rhythmic alterations of both CCM and PR transcript levels are unlinked to circadian clock: the "free-running state" reveals no discernible rhythmicity in transcript changes. Only continuous light leads to high transcript levels but no detectable transcripts were observed in continuous dark. Asynchronous continuous light cultures, upon shifting to low from high CO exhibit only transient induction of PR transcripts/proteins while CCM transcript induction is stable, indicating the loss of co-regulation between PR and CCM gene transcription. Lastly, we also describe that both CCM and PR transcripts/proteins are induced in low CO even in mixotrophic cultures, but only in high light, the same being attenuated in high CO, implying that high light is a mandatory "trigger" for CCM and PR induction in low CO mixotrophy. Our study provides comprehensive analyses of conditions where CCM and PR were differently regulated, setting a paradigm for a detailed mechanistic probing of these responses.
在莱茵衣藻中,碳浓缩机制(CCM)和光呼吸(PR)相互关联且共同调节,但共同调节发生改变的条件尚未得到充分探索。在此,我们发现,即使在黑暗中且两个过程均不活跃时,PR基因转录本也会像CCM转录本一样被诱导。这种昼夜循环表明,转录本水平在12小时光照的中间时段达到峰值,在12小时黑暗的早期下降,随后在黑暗中期再次开始上升。有趣的是,黑暗中期的起始对高浓度二氧化碳敏感,这意味着即使在黑暗中,活跃的碳传感机制也在运行。CCM和PR转录本水平的节律性变化与生物钟无关:“自由运行状态”在转录本变化中没有明显的节律性。只有持续光照会导致转录本水平升高,但在持续黑暗中未观察到可检测到的转录本。从高浓度二氧化碳转变为低浓度二氧化碳时,异步连续光照培养物仅短暂诱导PR转录本/蛋白质,而CCM转录本诱导则稳定,这表明PR和CCM基因转录之间的共同调节丧失。最后,我们还描述了即使在混合营养培养物中,低浓度二氧化碳也会诱导CCM和PR转录本/蛋白质,但仅在高光条件下,在高浓度二氧化碳中这种诱导会减弱,这意味着高光对于低浓度二氧化碳混合营养中CCM和PR的诱导是一个必要的“触发因素”。我们的研究对CCM和PR受到不同调节的条件进行了全面分析,为详细探究这些反应的机制树立了典范。