Wallner Thomas, Hagiwara Yoshinori, Bernát Gábor, Sobotka Roman, Reijerse Edward J, Frankenberg-Dinkel Nicole, Wilde Annegret
Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Heinrich-Buff-Ring 26-32, Germany.
Biochim Biophys Acta. 2012 Nov;1817(11):2016-26. doi: 10.1016/j.bbabio.2012.06.002. Epub 2012 Jun 7.
Ycf34 is a hypothetical chloroplast open reading frame that is present in the chloroplast genomes of several non-green algae. Ycf34 homologues are also encoded in all sequenced genomes of cyanobacteria. To evaluate the role of Ycf34 we have constructed and analysed a cyanobacterial mutant strain. Inactivation of ycf34 in Synechocystis sp. PCC 6803 showed no obvious phenotype under normal light intensity growth conditions. However, when the cells were grown under low light intensity they contained less and smaller phycobilisome antennae and showed a strongly retarded growth, suggesting an essential role of the Ycf34 polypeptide under light limiting conditions. Northern blot analysis revealed a very weak expression of the phycocyanin operon in the ycf34 mutant under light limiting growth in contrast to the wild type and to normal light conditions. Oxygen evolution and P(700) measurements showed impaired electron flow between photosystem II and photosystem I under these conditions which suggest that the impaired antenna size is most likely due to a highly reduced plastoquinone pool which triggers regulation on a transcriptional level. Using a FLAG-tagged Ycf34 we found that this protein is tightly bound to the thylakoid membranes. UV-vis and Mössbauer spectroscopy of the recombinant Ycf34 protein demonstrate the presence of an iron-sulphur cluster. Since Ycf34 lacks homology to known iron-sulphur cluster containing proteins, it might constitute a new type of iron-sulphur protein implicated in redox signalling or in optimising the photosynthetic electron transport chain.
Ycf34是一个假定的叶绿体开放阅读框,存在于几种非绿色藻类的叶绿体基因组中。蓝细菌的所有已测序基因组中也编码了Ycf34同源物。为了评估Ycf34的作用,我们构建并分析了一个蓝细菌突变株。集胞藻PCC 6803中ycf34的失活在正常光照强度生长条件下未表现出明显的表型。然而,当细胞在低光照强度下生长时,它们含有更少且更小的藻胆体天线,并表现出生长严重迟缓,这表明Ycf34多肽在光照限制条件下具有重要作用。Northern印迹分析显示,与野生型和正常光照条件相比,在光照限制生长条件下,ycf34突变体中藻蓝蛋白操纵子的表达非常弱。氧气释放和P(700)测量表明,在这些条件下,光系统II和光系统I之间的电子流受损,这表明天线尺寸受损很可能是由于质体醌库高度减少,从而触发了转录水平的调控。使用带有FLAG标签的Ycf34,我们发现该蛋白与类囊体膜紧密结合。重组Ycf34蛋白的紫外可见光谱和穆斯堡尔光谱证明存在铁硫簇。由于Ycf34与已知的含铁硫簇蛋白缺乏同源性,它可能构成一种新型的铁硫蛋白,参与氧化还原信号传导或优化光合电子传递链。