Aguilera Selene, Alvarez-Morales Ariel, Murillo Jesús, Hernández-Flores José Luis, Bravo Jaime, De la Torre-Zavala Susana
Laboratorio Integral de Investigación en Alimentos. CONACYT-Instituto Tecnológico de Tepic, Tepic, Nayarit, México.
Departamento de Ingeniería Genética, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Irapuato, Guanajuato, México.
PLoS One. 2017 Jun 1;12(6):e0178441. doi: 10.1371/journal.pone.0178441. eCollection 2017.
Pseudomonas syringae pv. phaseolicola produces phaseolotoxin in a temperature dependent manner, being optimally synthesized between 18°C and 20°C, while no detectable amounts are present above 28°C. The Pht cluster, involved in the biosynthesis of phaseolotoxin, contains 23 genes that are organized in five transcriptional units. The function of most of the genes from the Pht cluster is still unknown and little information about the regulatory circuitry leading to expression of these genes has been reported. The purpose of the present study was to investigate the participation of pht genes in the regulation of the operons coded into the Pht cluster. We conducted Northern blot, uidA fusions and reverse transcription-PCR assays of pht genes in several mutants unable to produce phaseolotoxin. This allowed us to determine that, in P. syringae pv. phaseolicola NPS3121, genes phtABC are essential to prevent their own expression at 28°C, a temperature at which no detectable amounts of the toxin are present. We obtained evidence that the phtABC genes also participate in the regulation of the phtD, phtM and phtL operons. According to our results, we propose that PhtABC and other Pht product activities could be involved in the synthesis of the sulfodiaminophosphinyl moiety of phaseolotoxin, which indirectly could be involved in the transcriptional regulation of the phtA operon.
菜豆假单胞菌菜豆致病变种以温度依赖的方式产生菜豆毒素,在18°C至20°C之间合成量最佳,而在28°C以上则不存在可检测到的量。参与菜豆毒素生物合成的Pht基因簇包含23个基因,这些基因被组织成五个转录单元。Pht基因簇中大多数基因的功能仍然未知,关于导致这些基因表达的调控电路的信息报道很少。本研究的目的是研究pht基因在调控Pht基因簇编码的操纵子中的作用。我们对几个无法产生菜豆毒素的突变体中的pht基因进行了Northern印迹、uidA融合和逆转录PCR分析。这使我们能够确定,在菜豆假单胞菌菜豆致病变种NPS3121中,基因phtABC对于在28°C(此时不存在可检测到的毒素量)时阻止其自身表达至关重要。我们获得的证据表明,phtABC基因也参与phtD、phtM和phtL操纵子的调控。根据我们的结果,我们提出PhtABC和其他Pht产物活性可能参与菜豆毒素磺基二氨基磷酰基部分的合成,这可能间接参与phtA操纵子的转录调控。