Department of Biological Sciences, Binghamton University, Binghamton, NY, 13902, USA.
Binghamton Biofilm Research Center, Binghamton University, Binghamton, NY, 13902, USA.
NPJ Biofilms Microbiomes. 2022 Jul 7;8(1):54. doi: 10.1038/s41522-022-00314-y.
Transcriptomic studies have revealed a large number of uncharacterized genes that are differentially expressed in biofilms, which may be important in regulating biofilm phenotypes such as resistance to antimicrobial agents. To identify biofilm genes of unknown function in P. aeruginosa, we made use of RNA-seq and selected 27 uncharacterized genes that were induced upon biofilm growth. Biofilms by respective mutants were subsequently analyzed for two biofilm characteristics, the biofilm architecture and drug susceptibility. The screen revealed 12 out of 27 genes to contribute to biofilm formation and 13 drug susceptibility, with 8 genes affecting both biofilm phenotypes. Amongst the genes affecting both biofilm phenotypes was PA2146, encoding a small hypothetical protein that exhibited some of the most substantial increases in transcript abundance during biofilm growth by P. aeruginosa PAO1 and clinical isolates. PA2146 is highly conserved in ɣ-proteobacteria. Inactivation of PA2146 affected both biofilm phenotypes in P. aeruginosa PAO1, with inactivation of homologs in Klebsiella pneumoniae and Escherichia coli having similar effects. Heterologous expression of PA2146 homologs complemented the P. aeruginosa ∆PA2146, suggesting that PA2146 homologs substitute for and play a similar role as PA2146 in P. aeruginosa.
转录组研究揭示了大量在生物膜中差异表达的未被描述的基因，这些基因可能在调节生物膜表型方面很重要，如对抗生素的耐药性。为了鉴定铜绿假单胞菌中未知功能的生物膜基因，我们利用 RNA-seq 技术选择了 27 个在生物膜生长时被诱导的未被描述的基因。随后，对相应突变体的生物膜进行了分析，以研究生物膜结构和药物敏感性两个特征。该筛选揭示了 27 个基因中的 12 个与生物膜形成有关，13 个与药物敏感性有关，其中 8 个基因同时影响这两种生物膜表型。在影响这两种生物膜表型的基因中，有编码一个小的假设蛋白的 PA2146 基因，该蛋白在铜绿假单胞菌 PAO1 和临床分离株的生物膜生长过程中表现出转录丰度的最大程度增加。PA2146 在γ-变形菌中高度保守。PA2146 的失活影响铜绿假单胞菌 PAO1 的两种生物膜表型，而肺炎克雷伯菌和大肠杆菌同源基因的失活也有类似的影响。PA2146 同源基因的异源表达补充了 P. aeruginosa ∆PA2146，表明 PA2146 同源基因在铜绿假单胞菌中替代并发挥类似的作用。
