The MarR family regulator RmaH mediates acid tolerance of Lactococcus lactis through regulating peptidoglycan modification genes

• Published in: Journal of dairy science 2024-12, Vol.107 (12), p.10383-10395
• Main Authors: Song, Qianqian, Wu, Hao, Zhang, Peng, Zhu, Hongji, Xie, Jiawei, Liu, Jiaheng, Qiao, Jianjun
• Format: Article
• Language: English
• Subjects: Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Gene Expression Regulation, Bacterial; lactic acid stress; Lactococcus lactis; Lactococcus lactis - genetics; Lactococcus lactis - metabolism; murT-gatD operon; Peptidoglycan - metabolism; transcription regulator RmaH
• ISSN: 0022-0302; 1525-3198; 1525-3198
• DOI: 10.3168/jds.2024-25152

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Lactococcus lactis, widely used in the food fermentation industry, has developed various ways to regulate acid adaptation in the process of evolution. Investigation into how peptidoglycan (PG) senses and responds to acid stress is an expanding field. Here, we addressed the regulation of murT-gatD genes, which are responsible for the amidation of PG d-Glu. We found that lactic acid stress reduced murT-gatD expression, and overexpressing these genes notably decreased acid tolerance of L. lactis NZ9000, possibly due to a reduction in the negative charge of PG, thereby facilitating the influx of extracellular protons into the cell. Subsequently, by using a combination of DNA pulldown assay and electrophoretic mobility shift assay, we identified a novel MarR family regulator, RmaH, as an activator of murT-gatD transcription. Further MEME motif prediction, electrophoretic mobility shift assay verification, and fluorescent protein reporter assay showed that RmaH directly bound to the DNA motif 5′-KGVAWWTTTTGCT-3′ located in the upstream region of murT-gatD. Beyond the mechanistic investigation of RmaH activation of murT-gatD, this study provides new insight into how PG modification is regulated and responds to lactic acid stress.