Project description:Gene expression in Lactococcus lactis MG1363 was compared to that of L. lactis MG1363 â??guaA in rich GM17 medium. One condition design comparison of two strains
Project description:Lactococcus lactis NZ9000 and its parent MG1363 are the most commonly used lactic acid bacteria for expression and physiological studies. We noted unexpected but significant differences in the growth behaviors of both strains. We sequenced the entire genomes of the original NZ9000 and MG1363 strains using an ultradeep sequencing strategy. The analysis of the L. lactis NZ9000 genome yielded 79 differences, mostly point mutations, with the annotated genome sequence of L. lactis MG1363. Resequencing of the MG1363 strain revealed that 73 out of the 79 differences were due to errors in the published sequence. Comparative transcriptomic studies revealed several differences in the regulation of genes involved in sugar fermentation, which can be explained by two specific mutations in a region of the ptcC promoter with a key role in the regulation of cellobiose and glucose uptake.
Project description:The contribution of small RNAs to the regulation of cell wall defense mechanisms, which play a central role in protecting bacteria from environmental insults, remains poorly understood in Lactococcus cremoris, a paradigmatic organism for studying lactic acid bacteria. Analysis of the profiles of 193 putative sRNAs, including 131 newly identified in this study, revealed altered expression of 20 and 25 sRNAs following treatment with major cell wall-targeting antimicrobials lysozyme and penicillin G, respectively. Further analysis and genome-wide functional screening identified four sRNAs that modulate the lysozyme response in opposite directions, with sLLM1042+, sLLM2-, and sLLM1993+ conferring increased, and sLLM461+ decreased, resistance. Moreover, elevated expression of sLLM2- or sLLM1042+ promoted the sensitivity of cells to penicillin G and certain abiotic stresses, and had pleiotropic effects on gene regulation at both RNA and protein levels, positioning them as key elements in regulatory networks. Notably, the sLLM2- mediated decrease in four putative tellurium resistance proteins resulted in L. cremoris sensitivity to tellurite stress. As sLLM2- mutations, altering lysozyme and penicillin responses, reduce the expression of alanine racemase Alr, we suggest that its upregulation possibly modulates D-alanine levels, thereby affecting lipoteichoic acid modification or peptidoglycan synthesis. Our findings underscored the broad roles of sRNAs in defensive responses to cell wall-targeting antimicrobials in lactic acid bacteria.