Liu Y, Fletcher H M
Department of Microbiology and Molecular Genetics, School of Medicine, Loma Linda University, Loma Linda, California 92350, USA.
Oral Microbiol Immunol. 2001 Jun;16(3):136-43. doi: 10.1034/j.1399-302x.2001.016003136.x.
The recA gene product in Porphyromonas gingivalis is involved in DNA repair. Further, disruption of this gene can affect the proteolytic activity and expression of other virulence factors in this organism. Since several known environmental factors can influence virulence gene expression in P. gingivalis, we investigated the influence of these signals on the expression of the recA gene in this organism. A heterodiploid strain of P. gingivalis (designated FLL118) containing a transcriptional fusion of the recA promoter region and the promoterless tetracycline-resistant gene [tetA(Q)2] and xylosidase/arabinosidase (xa) gene cassette was constructed. The recA promoter activity was assessed by measurement of xylosidase activity in FLL118. The expression remained relatively constant during different growth phases, at different pH levels and in the presence of DNA-damaging agents. In response to hemin limitation and in the presence of calcium there was a moderate increase in recA promoter activity. Temperature also affected the expression. The highest level of xylosidase activity was observed in cultures at 32 degrees C with a decline of approximately 46% as growth temperature increased to 41 degrees C. Reverse transcriptase polymerase chain reaction analysis revealed that this regulation may be occurring at the transcriptional level. These results suggest that expression of the recA gene in P. gingivalis W83 is responsive to several environmental signals but is not regulated by a DNA damage-inducible SOS-like regulatory system.
牙龈卟啉单胞菌中的recA基因产物参与DNA修复。此外,该基因的破坏会影响该生物体中其他毒力因子的蛋白水解活性和表达。由于几种已知的环境因素可影响牙龈卟啉单胞菌中毒力基因的表达,因此我们研究了这些信号对该生物体中recA基因表达的影响。构建了一种牙龈卟啉单胞菌的异源二倍体菌株(命名为FLL118),其包含recA启动子区域与无启动子的四环素抗性基因[tetA(Q)2]和木糖苷酶/阿拉伯糖苷酶(xa)基因盒的转录融合体。通过测量FLL118中的木糖苷酶活性来评估recA启动子活性。在不同生长阶段、不同pH水平以及存在DNA损伤剂的情况下,该表达保持相对恒定。在血红素限制和存在钙的情况下,recA启动子活性有适度增加。温度也影响表达。在32℃培养物中观察到最高水平的木糖苷酶活性,随着生长温度升至41℃,其活性下降约46%。逆转录酶聚合酶链反应分析表明,这种调节可能发生在转录水平。这些结果表明,牙龈卟啉单胞菌W83中recA基因的表达对几种环境信号有反应,但不受DNA损伤诱导的SOS样调节系统的调控。