Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, Corvallis, Oregon, United States of America.
PLoS One. 2013 May 13;8(5):e63336. doi: 10.1371/journal.pone.0063336. Print 2013.
We previously found that inhibition of the TCA cycle, either through mutations or chemical inhibition, increased toxT transcription in Vibrio cholerae. In this study, we found that the addition of malonate, an inhibitor of succinate dehydrogenase (SDH), decreased toxT transcription in V. cholerae, an observation inconsistent with the previous pattern observed. Unlike another SDH inhibitor, 2-thenoyltrifluoroacetone (TTFA), which increased toxT transcription and slightly inhibited V. cholerae growth, malonate inhibited toxT transcription in both the wild-type strain and TCA cycle mutants, suggesting malonate-mediated inhibition of virulence gene expression is independent to TCA cycle activity. Addition of malonate also inhibited ctxB and tcpA expressions but did not affect aphA, aphB, tcpP and toxR expressions. Malonate inhibited cholera toxin (CT) production in both V. cholerae classical biotype strains O395N1 and CA401, and El Tor biotype strain, N16961. Consistent with previous reports, we confirmed that these strains of V. cholerae did not utilize malonate as a primary carbon source. However, we found that the addition of malonate to the growth medium stimulated V. cholerae growth. All together, these results suggest that metabolizing malonate as a nutrient source negatively affects virulence gene expression in V. cholerae.
我们之前发现,无论是通过突变还是化学抑制,三羧酸循环的抑制都会增加霍乱弧菌中 toxT 的转录。在这项研究中,我们发现丙二酸盐(琥珀酸脱氢酶 SDH 的抑制剂)的添加降低了霍乱弧菌中的 toxT 转录,这一观察结果与之前观察到的模式不一致。与另一种 SDH 抑制剂 2-(三氟乙酰基)苯甲酰氟(TTFA)不同,TTFA 增加 toxT 转录并轻微抑制霍乱弧菌生长,丙二酸盐在野生型菌株和 TCA 循环突变体中均抑制 toxT 转录,表明丙二酸盐介导的毒力基因表达抑制与 TCA 循环活性无关。丙二酸盐的添加也抑制了 ctxB 和 tcpA 的表达,但不影响 aphA、aphB、tcpP 和 toxR 的表达。丙二酸盐抑制了霍乱毒素(CT)在霍乱弧菌经典生物型菌株 O395N1 和 CA401 以及 El Tor 生物型菌株 N16961 中的产生。与之前的报道一致,我们证实这些霍乱弧菌菌株不将丙二酸盐用作主要碳源。然而,我们发现向生长培养基中添加丙二酸盐刺激了霍乱弧菌的生长。总的来说,这些结果表明,将丙二酸盐作为营养源代谢会对霍乱弧菌中的毒力基因表达产生负面影响。