Kiss Eva, Kós Péter B, Vass Imre
Institute of Plant Biology, Biological Research Center, Temesvari krt. 62, 6726 Szeged, Hungary.
J Biotechnol. 2009 Jun 1;142(1):31-7. doi: 10.1016/j.jbiotec.2009.02.007. Epub 2009 Feb 20.
To identify optimal conditions for renewable hydrogen production from sunlight and water we have studied transcriptional changes of the hoxEFUYH genes encoding the bidirectional hydrogenase in the cyanobacterium Synechocystis PCC 6803. Transcript abundance detection by real time polymerase chain reaction was supplemented with variable chlorophyll fluorescence measurements to monitor redox changes of the photosynthetic electron transport chain. Our main observations are: (i) abundance of hox transcripts decreases in the dark and recovers in the light. (ii) Inhibition of the Calvin cycle by glycolaldehyde suppresses hox gene transcription, which can be restored by the addition of electron transport inhibitors 3-(3,4-dichlorophenyl)-1,1-dimethylurea and dibromothymoquinone. (iii) The transcript levels of all hox genes are increased in anoxia, with additional induction of hoxEF in darkness or in the presence of dibromothymoquinone. (iv) Plastoquinone pool redox changes are not correlated with hox transcript level changes. (v) Changes in the transcript levels of lexA and sll0359 genes, encoding putative regulators of hox genes, are only partly correlated with transcript changes of hox genes under different conditions. Our data demonstrate a previously unrecognized light- and oxygen-dependent regulation of hox gene transcription in Synechocystis PCC 6803, which is related to photosynthetic electron transport and to unidentified oxygen and redox sensors. We also conclude that neither LexA nor Sll0359 are likely to be exclusive regulators of hox gene transcription.
为了确定利用阳光和水进行可再生制氢的最佳条件,我们研究了编码蓝藻集胞藻PCC 6803中双向氢化酶的hoxEFUYH基因的转录变化。通过实时聚合酶链反应检测转录本丰度,并辅以可变叶绿素荧光测量,以监测光合电子传递链的氧化还原变化。我们的主要观察结果如下:(i) hox转录本丰度在黑暗中降低,在光照下恢复。(ii) 乙醇醛对卡尔文循环的抑制作用会抑制hox基因转录,添加电子传递抑制剂3-(3,4-二氯苯基)-1,1-二甲基脲和二溴百里香醌可恢复这种抑制作用。(iii) 在缺氧条件下,所有hox基因的转录水平都会升高,在黑暗中或存在二溴百里香醌的情况下,hoxEF会额外诱导表达。(iv) 质体醌库的氧化还原变化与hox转录本水平变化不相关。(v) 编码hox基因假定调节因子的lexA和sll0359基因的转录水平变化,在不同条件下仅部分与hox基因的转录变化相关。我们的数据表明,集胞藻PCC 6803中hox基因转录存在以前未被认识的光和氧依赖性调节,这与光合电子传递以及未知的氧和氧化还原传感器有关。我们还得出结论,LexA和Sll0359都不太可能是hox基因转录的唯一调节因子。