Jones Sherry Usun, Kee Boon Pin, Chew Ching Hoong, Yeo Chew Chieng, Chua Kek Heng, Puah Suat Moi
Department of Biomedical Science, Faculty of Medicine, Universiti Malaya, 50603, Kuala Lumpur, Malaysia.
Faculty of Health Sciences, Universiti Sultan Zainal Abidin, 21300, Kuala Nerus, Terengganu, Malaysia.
Heliyon. 2024 Oct 19;10(20):e39634. doi: 10.1016/j.heliyon.2024.e39634. eCollection 2024 Oct 30.
Bacterial small RNAs (sRNAs) play crucial roles in coordinating gene regulatory networks in various physiological processes, including biofilm formation. In this study, RNA sequencing was performed on biofilm ( = 4) and planktonic ( = 4) cells harvested at 10 h (pre-stationary phase of biofilm development) to identify biofilm-associated sRNAs in human methicillin-susceptible (MSSA) recovered from urine isolate. A total of 56 highly expressed sRNAs were identified with 15 overlapping sRNA genes (srn_9348, sprD, sRNA205, sRNA288, srn_2467, Sau-25, srn_2468, sRNA260, sRNA200, RsaE, sRNA397, Teg55, Teg60, RsaX05 and Teg140). Further validation through RT-qPCR analysis of nine sRNAs revealed that srn_9348 and sRNA260 were significantly expressed in the biofilm cells of urine sample. Both sRNAs were predicted to interact with mRNA genes including intracellular adhesin A () and host factor protein () involved in biofilm formation via -acting and -acting using CopraRNA analysis. Therefore, both sRNAs merit further investigations via reverse genetic approaches to elucidate their mechanism of translational regulation. In summary, the transcriptomic analysis conducted in this study offers new insights into the potential regulatory roles of sRNAs in MSSA biofilm development within the urinary environment.
细菌小RNA（sRNA）在协调包括生物膜形成在内的各种生理过程中的基因调控网络中发挥着关键作用。在本研究中，对在10小时（生物膜发育的静止前期）收获的生物膜（n = 4）和浮游（n = 4）细胞进行了RNA测序，以鉴定从尿液分离物中获得的人甲氧西林敏感金黄色葡萄球菌（MSSA）中与生物膜相关的sRNA。共鉴定出56个高表达的sRNA，其中有15个重叠的sRNA基因（srn_9348、sprD、sRNA205、sRNA288、srn_2467、Sau-25、srn_2468、sRNA260、sRNA200、RsaE、sRNA397、Teg55、Teg60、RsaX05和Teg140）。通过对9个sRNA的RT-qPCR分析进一步验证发现，srn_9348和sRNA260在尿液样本的生物膜细胞中显著表达。使用CopraRNA分析预测这两个sRNA均与包括参与生物膜形成的细胞内黏附素A（icaA）和宿主因子蛋白（hfp）在内的mRNA基因通过反式作用和顺式作用相互作用。因此，这两个sRNA都值得通过反向遗传学方法进行进一步研究，以阐明其翻译调控机制。总之，本研究进行的转录组分析为sRNA在泌尿环境中MSSA生物膜发育中的潜在调控作用提供了新的见解。
