Identification of c-di-GMP/FleQ-Regulated New Target Genes, Including cyaA, Encoding Adenylate Cyclase, in Pseudomonas putida.
Identification of c-di-GMP/FleQ-Regulated New Target Genes, Including cyaA, Encoding Adenylate Cyclase, in Pseudomonas putida.
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恶臭假单胞菌中 c-di-GMP/FleQ 调控的新靶基因的鉴定，包括编码腺苷酸环化酶的 cyaA
DOI：
10.1128/msystems.00295-21
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发表时间：
2021-05-11
期刊：
影响因子：
6.4
通讯作者：
Huang Q
中科院分区：
文献类型：
--
作者：
Xiao Y;Chen H;Nie L;He M;Peng Q;Zhu W;Nie H;Chen W;Huang Q
c-di-GMP/FleQ在转录水平上通过FleQ促进假单胞菌从浮游生物到生物膜的生活方式转变。发现c-di-GMP/FleQ直接调控的新靶基因有助于拓宽对c-di-GMP/FleQ介导的转录调控的认识。细菌第二信使环双胍酸酯（c-di-GMP）通过其转录调控效应物FleQ调节假单胞菌物种从浮游生物到生物膜的生活方式转变。FleQ调节生物膜和鞭毛相关基因的转录，以响应c-di-GMP。通过转录组学分析和FleQ- dna结合实验，本研究在p.p putida中发现了5个新的c-di-GMP/FleQ靶基因，分别是PP_0681、PP_0788、PP_4519 （lapE）、PP_5222 （cyaA）和PP_5586。除了编码外膜孔蛋白的lapE和编码腺苷酸环化酶的cyaA外，其他三个编码假想蛋白的基因的功能尚不清楚。FleQ和c-di-GMP协同抑制PP_0788和cyaA的转录，促进PP_0681、lapE和PP_5586的转录。体外和体内实验均表明FleQ可直接与这五个基因的启动子结合。进一步分析证实，LapE在粘附素LapA的分泌中起核心作用，c-di-GMP/FleQ增加LapE的转录，从而促进粘附素分泌和生物膜的形成。腺苷酸环化酶CyaA负责另一个第二信使环AMP （cAMP）的合成。FleQ与c-di-GMP协同降低cAMP含量，提示c-di-GMP与FleQ通过调节cyaA的表达共同调控cAMP。总的来说，本研究为c-di-GMP/FleQ调控基因家族增加了5个新成员，揭示了c-di-GMP/FleQ在恶臭假单胞菌LapA分泌和cAMP合成调控中的作用。c-di-GMP/FleQ在转录水平上通过FleQ促进假单胞菌从浮游生物到生物膜的生活方式转变。发现c-di-GMP/FleQ直接调控的新靶基因有助于拓宽对c-di-GMP/FleQ介导的转录调控的认识。c-di-GMP/FleQ对LapA的调控保证了高效的LapA分泌和生物膜的形成。P. putida和P. aeruginosa中c-di-GMP与cAMP负相关的机制尚不清楚。我们关于c-di-GMP/FleQ对cyaA转录抑制的研究结果揭示了c-di-GMP降低p.p putida cAMP含量的机制。
c-di-GMP/FleQ promotes the plankton-to-biofilm lifestyle transition at the transcriptional level via FleQ in Pseudomonas species. Identification of new target genes directly regulated by c-di-GMP/FleQ helps to broaden the knowledge of c-di-GMP/FleQ-mediated transcriptional regulation. The bacterial second messenger cyclic diguanylate (c-di-GMP) modulates plankton-to-biofilm lifestyle transition of Pseudomonas species through its transcriptional regulatory effector FleQ. FleQ regulates transcription of biofilm- and flagellum-related genes in response to c-di-GMP. Through transcriptomic analysis and FleQ-DNA binding assay, this study identified five new target genes of c-di-GMP/FleQ in P. putida, including PP_0681, PP_0788, PP_4519 (lapE), PP_5222 (cyaA), and PP_5586. Except lapE encoding an outer membrane pore protein and cyaA encoding an adenylate cyclase, the functions of the other three genes encoding hypothetical proteins remain unknown. FleQ and c-di-GMP coordinately inhibit transcription of PP_0788 and cyaA and promote transcription of PP_0681, lapE, and PP_5586. Both in vitro and in vivo assays show that FleQ binds directly to promoters of the five genes. Further analyses confirm that LapE plays a central role of in the secretion of adhesin LapA and that c-di-GMP/FleQ increases lapE transcription, thereby promoting adhesin secretion and biofilm formation. The adenylate cyclase CyaA is responsible for synthesis of another second messenger, cyclic AMP (cAMP). FleQ and c-di-GMP coordinate to decrease the content of cAMP, suggesting that c-di-GMP and FleQ coregulate cAMP by modulating cyaA expression. Overall, this study adds five new members to the c-di-GMP/FleQ-regulated gene family and reveals the role of c-di-GMP/FleQ in LapA secretion and cAMP synthesis regulation in P. putida. IMPORTANCE c-di-GMP/FleQ promotes the plankton-to-biofilm lifestyle transition at the transcriptional level via FleQ in Pseudomonas species. Identification of new target genes directly regulated by c-di-GMP/FleQ helps to broaden the knowledge of c-di-GMP/FleQ-mediated transcriptional regulation. Regulation of lapE by c-di-GMP/FleQ guarantees highly efficient LapA secretion and biofilm formation. The mechanism of negative correlation between c-di-GMP and cAMP in both P. putida and P. aeruginosa remains unknown. Our result concerning transcriptional inhibition of cyaA by c-di-GMP/FleQ reveals the mechanism underlying the decrease of cAMP content by c-di-GMP in P. putida.