Inouye Takayuki, Odahara Masaki, Fujita Tomomichi, Hasebe Mitsuyasu, Sekine Yasuhiko
Department of Life Science, College of Science, Rikkyo (St. Paul's) University, Tokyo, Japan.
Biosci Biotechnol Biochem. 2008 May;72(5):1340-7. doi: 10.1271/bbb.80014. Epub 2008 May 7.
RecA protein is widespread in bacteria, and it plays a crucial role in homologous recombination. We have identified two bacterial-type recA gene homologs (PprecA1, PprecA2) in the cDNA library of the moss Physcomitrella patens. N-terminal fusion of the putative organellar targeting sequence of PpRecA2 to the green fluorescent protein (GFP) caused a targeting of PpRecA2 to the chloroplasts. Mutational analysis showed that the first AUG codon acts as initiation codon. Fusion of the full-length PpRecA2 to GFP caused the formation of foci that were colocalized with chloroplast nucleoids. The amounts of PprecA2 mRNA and protein in the cells were increased by treatment with DNA damaging agents. PprecA2 partially complemented the recA mutation in Escherichia coli. These results suggest the involvement of PpRecA2 in the repair of chloroplast DNA.
RecA蛋白在细菌中广泛存在,并且在同源重组中发挥关键作用。我们在小立碗藓的cDNA文库中鉴定出了两个细菌型recA基因同源物(PprecA1、PprecA2)。将PpRecA2推测的细胞器靶向序列与绿色荧光蛋白(GFP)进行N端融合,导致PpRecA2靶向叶绿体。突变分析表明,第一个AUG密码子作为起始密码子。将全长PpRecA2与GFP融合导致形成与叶绿体类核共定位的聚焦点。用DNA损伤剂处理后,细胞中PprecA2 mRNA和蛋白质的量增加。PprecA2部分互补大肠杆菌中的recA突变。这些结果表明PpRecA2参与叶绿体DNA的修复。