Identification and characterisation of G-quadruplex DNA-forming sequences in the Pseudomonas aeruginosa genome.
Identification and characterisation of G-quadruplex DNA-forming sequences in the Pseudomonas aeruginosa genome.
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DOI：
10.1039/d2cb00205a
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发表时间：
2023-01-04
影响因子：
4.1
通讯作者：
Vilar, Ramon
中科院分区：
文献类型：
--
作者：
Evans, Lindsay;Kotar, Anita;Valentini, Martina;Filloux, Alain;Jamshidi, Shirin;Plavec, Janez;Rahman, Khondaker Miraz;Vilar, Ramon
一些革兰氏阴性菌，如铜绿假单胞菌，对一线抗生素产生耐药性。因此，迫切需要找到用于开发新药的替代生物分子靶标。由于非规范的DNA结构，如鸟嘌呤四链体（G4）已涉及在调节转录，我们有兴趣在确定是否有推定的四链体形成序列（PQS）在铜绿假单胞菌的基因组。使用生物信息学工具，我们筛选了36个可能与PQS存在的耐药性相关的基因，并选择其中10个进行生物物理表征（即圆二色性和热差UV/维斯光谱）。这些研究表明，这些富含G的序列（连接到murE，ftsB和mexC基因）形成稳定的鸟嘌呤四链体，通过NMR光谱研究;其中一个序列（mexC）的详细分析证实，它在K+离子的存在下采用了两个四重反平行四链体结构。我们还表明，FRET熔解分析，小分子可以稳定这三个新的G4 DNA结构在生理条件下。这些初步结果可能对开发具有替代生物分子靶点的新抗生素具有未来的意义，这反过来将有助于解决抗生素耐药性问题。我们报告了铜绿假单胞菌基因组中与抗菌素耐药性相关的基因中G-四链体DNA形成序列的鉴定和特征。
A number of Gram-negative bacteria such as Pseudomonas aeruginosa are becoming resistant to front-line antibiotics. Consequently, there is a pressing need to find alternative bio-molecular targets for the development of new drugs. Since non-canonical DNA structures such as guanine-quadruplexes (G4s) have been implicated in regulating transcription, we were interested in determining whether there are putative quadruplex-forming sequences (PQS) in the genome of Pseudomonas aeruginosa. Using bioinformatic tools, we screened 36 genes potentially relevant to drug resistance for the presence of PQS and 10 of these were selected for biophysical characterisation (i.e. circular dichroism and thermal difference UV/Vis spectroscopy). These studies showed that three of these G-rich sequences (linked to murE, ftsB and mexC genes) form stable guanine-quadruplexes which were studied by NMR spectroscopy; detailed analysis of one of the sequences (mexC) confirmed that it adopts a two-quartet antiparallel quadruplex structure in the presence of K+ ions. We also show by FRET melting assays that small molecules can stabilise these three new G4 DNA structures under physiological conditions. These initial results could be of future interest in the development of new antibiotics with alternative bio-molecular targets which in turn would help tackle antimicrobial resistance. We report identification and characterisation of G-quadruplex DNA forming sequences in the Pseudomonas aeruginosa genome in genes associated with antimicrobial resistance.
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影响因子：
14.9
作者：
Mergny JL;Li J;Lacroix L;Amrane S;Chaires JB
通讯作者：
Chaires JB
影响因子：
6.7
作者：
Harris LM;Merrick CJ
通讯作者：
Merrick CJ
影响因子：
21.8
作者：
Di Antonio M;Ponjavic A;Radzevičius A;Ranasinghe RT;Catalano M;Zhang X;Shen J;Needham LM;Lee SF;Klenerman D;Balasubramanian S
通讯作者：
Balasubramanian S
影响因子：
4.6
作者：
Bartas, Martin;Cutova, Michaela;Pecinka, Petr
通讯作者：
Pecinka, Petr
影响因子：
5
作者：
Kota, Swathi;Dhamodharan, V.;Misra, Hari S.
通讯作者：
Misra, Hari S.