Tamegai Hideyuki, Kawano Hiroaki, Ishii Akihiro, Chikuma Sayaka, Nakasone Kaoru, Kato Chiaki
Department of Chemistry, College of Humanities and Sciences, Nihon University, 3-25-40 Sakurajosui, Setagaya-ku, Tokyo 156-8550, Japan.
Extremophiles. 2005 Jun;9(3):247-53. doi: 10.1007/s00792-005-0439-2. Epub 2005 Apr 21.
The genes of cytochrome bd-encoding cydAB were identified from a deep-sea bacterium Shewanella violacea DSS12. These showed significant homologies with known cydAB gene sequences from various organisms. Additionally, highly conserved regions that are important for the enzymatic function were also conserved in cydA of S. violacea. Based on the results, transcriptional analysis of cydAB operon and cydDC operon (required for assembly of cytochrome bd) of S. violacea in microaerobic condition was performed under the growth condition of various pressures. The gene of cydA was expressed even under the condition of atmospheric pressure and its expression was enhanced with pressurization. On the other hand, the expression of cydC was strongly depressed under the condition of atmospheric pressure compared with the case under high pressure. It appeared spectrophotometrically that loss of cytochrome bd in S. violacea under atmospheric pressure shown in previous study is caused mainly by the loss of cydDC. Further, under the growth condition of atmospheric pressure, either less amount or no d-type cytochrome was expressed compared with the case of high-pressure condition even if the organism was grown under alkaline condition or in the presence of uncoupler, which are the inducible condition of d-type cytochrome in Escherichia coli. These results suggested that the significant amount of d-type cytochrome expression is specific event under the growth condition of high pressure.
从深海细菌紫色希瓦氏菌DSS12中鉴定出编码细胞色素bd的cydAB基因。这些基因与来自各种生物体的已知cydAB基因序列具有显著同源性。此外,对酶功能重要的高度保守区域在紫色希瓦氏菌的cydA中也保守。基于这些结果,在不同压力的生长条件下,对紫色希瓦氏菌在微需氧条件下的cydAB操纵子和cydDC操纵子(细胞色素bd组装所需)进行了转录分析。即使在大气压条件下,cydA基因也表达,并且其表达随着压力增加而增强。另一方面,与高压条件下相比,在大气压条件下cydC的表达强烈受抑制。分光光度法显示,先前研究中紫色希瓦氏菌在大气压下细胞色素bd的丧失主要是由cydDC的丧失引起的。此外,在大气压生长条件下,即使该生物体在碱性条件下生长或在解偶联剂存在下生长(这是大肠杆菌中d型细胞色素的诱导条件),与高压条件相比,d型细胞色素的表达量也较少或不表达。这些结果表明,大量d型细胞色素的表达是高压生长条件下的特定事件。