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Light Response of Pseudomonas putida KT2440 Mediated by Class II LitR, a Photosensor Homolog.
Journal of Bacteriology ( IF 3.2 ) Pub Date : 2020-09-23 , DOI: 10.1128/jb.00146-20
Satoru Sumi 1 , Naotaka Mutaguchi 1 , Teppei Ebuchi 1 , Hiroaki Tsuchida 1 , Takahiro Yamamoto 1 , Maki Suzuki 1 , Chihiro Natsuka 1 , Hatsumi Shiratori-Takano 1 , Masaki Shintani 2 , Hideaki Nojiri 3, 4 , Kenji Ueda 1 , Hideaki Takano 5
Affiliation  

Pseudomonas putida KT2440 retains three homologs (PplR1 to PplR3) of the LitR/CarH family, an adenosyl B12-dependent light-sensitive MerR family transcriptional regulator. Transcriptome analysis revealed the existence of a number of photoinducible genes, including pplR1, phrB (encoding DNA photolyase), ufaM (furan-containing fatty acid synthase), folE (GTP cyclohydrolase I), cryB (cryptochrome-like protein), and multiple genes without annotated/known function. Transcriptional analysis by quantitative reverse transcription-PCR with knockout mutants of pplR1 to pplR3 showed that a triple knockout completely abolished the light-inducible transcription in P. putida, which indicates the occurrence of ternary regulation of PplR proteins. A DNase I footprint assay showed that PplR1 protein specifically binds to the promoter regions of light-inducible genes, suggesting a consensus PplR1-binding direct repeat, 5′-T(G/A)TACAN12TGTA(C/T)A-3′. The disruption of B12 biosynthesis cluster did not affect the light-inducible transcription; however, disruption of ppSB1-LOV (where LOV indicates “light, oxygen, or voltage”) and ppSB2-LOV, encoding blue light photoreceptors adjacently located to pplR3 and pplR2, respectively, led to the complete loss of light-inducible transcription. Overall, the results suggest that the three PplRs and two PpSB-LOVs cooperatively regulate the light-inducible gene expression. The wide distribution of the pplR/ppSB-LOV cognate pair homologs in Pseudomonas spp. and related bacteria suggests that the response and adaptation to light are similarly regulated in the group of nonphototrophic bacteria.

中文翻译:

II类LitR(光传感器同源物)介导的恶臭假单胞菌KT2440的光响应。

恶臭假单胞菌KT2440保留了LitR / CarH家族的三个同源物(PplR1至PplR3),这是腺苷B 12依赖的光敏MerR家族的转录调节子。转录组分析揭示了许多光诱导性基因的存在,包括pplR1phrB(编码DNA光解酶),ufaM(含呋喃的脂肪酸合酶),folE(GTP环水解酶I),cryB(隐样蛋白)和多个基因没有带注释/已知功能。pplR1pplR3基因敲除突变体的定量逆转录PCR转录分析结果表明,三次敲除完全消除了恶臭假单胞菌的光诱导转录,这表明发生了PplR蛋白的三重调控。DNase I足迹测定法显示PplR1蛋白与光诱导基因的启动子区域特异性结合,提示共有PplR1结合直接重复序列5'-T(G / A)TACAN 12 TGTA(C / T)A-3 '。B 12生物合成簇的破坏不影响光诱导的转录。但是,ppSB1-LOV(其中LOV表示“光,氧或电压”)和ppSB2-LOV的破坏,编码了与pplR3pplR2相邻的蓝光感光体分别导致光诱导转录的完全丧失。总体而言,结果表明三个PplR和两个PpSB-LOV协同调节光诱导基因的表达。pplR / ppSB-LOV同源对同源物在假单胞菌中的广泛分布。相关细菌表明,在非光养细菌群中,对光的反应和适应性受到类似的调节。
更新日期:2020-09-23
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