Institut de Biologie et de Technologie de Saclay, Centre National de la Recherche Scientifique and Commissariat à l'Energie Atomique, 91191 Gif-sur-Yvette.
Mol Microbiol. 2011 Sep;81(5):1178-89. doi: 10.1111/j.1365-2958.2011.07739.x. Epub 2011 Jul 26.
Ferredoxin:NADP oxidoreductases (FNRs) constitute a family of flavoenzymes that catalyse the exchange of electrons between ferredoxin and NADP(H). In cyanobacteria FNR provides NADPH for photoautotrophic metabolism, but the enzyme is also capable of oxidizing NADPH providing reduced ferredoxin. In the cyanobacterium Synechocystis sp. strain PCC6803, the unique petH gene has two translation products depending on growth conditions. As a consequence two isoforms of the FNR accumulate - FNR(L) and FNR(S) . In the present work, analysis of petH expression reveals that different transcriptional start points (tsp) are responsible for this differential translation initiation. Under standard conditions (where FNR(L) accumulates), two tsps were found at -52 and -34 relative to the first translation start site. Under nitrogen-starvation conditions (where FNR(S) accumulates) a tsp was mapped at -126 relative to the first translation start site. Therefore, the transcript responsible for FNR(S) translation is longer than that producing FNR(L) . In addition, expression of the short or long transcript in E. coli resulted in the accumulation of FNR(L) or FNR(S) respectively. This result demonstrates that translation can initiate at two different sites, 336-bases apart (ATG-1 to ATG-113), depending only on the 5'UTR structure.
NADP 氧化还原酶(FNRs)构成了一类黄素酶,能够在铁氧还蛋白和 NADP(H)之间催化电子交换。在蓝藻中,FNR 为光合作用代谢提供 NADPH,但该酶也能够氧化 NADPH 提供还原型铁氧还蛋白。在集胞藻 PCC6803 中,独特的 petH 基因根据生长条件有两种翻译产物。因此,两种 FNR 同工酶积累——FNR(L)和 FNR(S)。在本工作中,对 petH 表达的分析表明,这种差异翻译起始是由不同的转录起始点(tsp)负责的。在标准条件下(积累 FNR(L)的情况下),在相对于第一个翻译起始位点 -52 和 -34 处发现了两个 tsp。在氮饥饿条件下(积累 FNR(S)的情况下),相对于第一个翻译起始位点,在 -126 处映射到一个 tsp。因此,负责 FNR(S)翻译的转录本比产生 FNR(L)的转录本更长。此外,在大肠杆菌中表达短或长的转录本分别导致 FNR(L)或 FNR(S)的积累。这一结果表明,翻译只能根据 5'UTR 结构从两个不同的位点起始,相隔 336 个碱基(ATG-1 到 ATG-113)。