Transcriptomics Analysis Uncovers Transient Ceftazidime Tolerance in Burkholderia Biofilms

生物膜 类鼻疽伯克霍尔德菌 微生物学 生物 头孢他啶 基因 转录组 多药耐受 基因表达 类鼻疽 细菌 遗传学 铜绿假单胞菌
作者
Supaksorn Chattagul,Mohd M. Khan,Alison Scott,Aleksandra Nita‐Lazar,Robert K. Ernst,David R. Goodlett,Rasana W. Sermswan
出处
期刊:ACS Infectious Diseases [American Chemical Society]
卷期号:7 (8): 2324-2336 被引量:3
标识
DOI:10.1021/acsinfecdis.1c00003
摘要

Burkholderia pseudomallei is an etiological agent of melioidosis, a severe community-acquired infectious disease. B. pseudomallei strain K96243 is sensitive to the drug ceftazidime (CAZ), but has been shown to exhibit transient CAZ tolerance when in a biofilm form. To investigate an observed shift in gene expression profile during CAZ tolerance condition and to better understand the mechanistic aspects of this transient tolerance, RNA-sequencing was performed on B. pseudomallei K96243 from the following three states: planktonic, biofilm, and planktonic shedding. Results indicated that the expression of 651 genes (10.97%) were significantly changed in both biofilm (resistant) and planktonic shedding (sensitive) cells in comparison to the planktonic state. The top four highly expressed genes identified in both states are associated with nitrosative stress response (BPSL2368), Fe–S homeostasis (BPSL2369), and nitrate respiration (BPSS1154 and BPSS1158). Additionally, five orthologous genes, BPSL2370–BPSL2374, implicated in Fe–S cluster biogenesis, and another gene, BPSL2863, involved in DNA-binding of the stress protein ferritin, were shown to increase expression by RT-qPCR. The shift in gene expression was especially prominent at the late stages of biofilm growth (72 and 96 h), specifically in the biofilm-challenged CAZ survivor cells. This suggested that in response to stress in a biofilm, differential expression of these genes may support development of the CAZ tolerance in Burkholderia. The application of iron chelator deferoxamine (DFO) to the biofilm caused a significant reduction in biofilm formation and associated CAZ tolerance. Therefore, the shift in Fe–S metabolism when B. pseudomallei is in a biofilm may help stabilize the levels of reactive oxygen species (ROS), thereby limiting tolerance to CAZ.
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