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A Novel LysR Family Factor STM0859 is Associated with The Responses of Salmonella Typhimurium to Environmental Stress and Biofilm Formation

ZHONGMEI MA
ZHONGMEI MA
, 
NA LI
NA LI
, 
CHENGCHENG NING
CHENGCHENG NING
, 
YUCHENG LIU
YUCHENG LIU
, 
YUN GUO
YUN GUO
, 
CHUNHUI JI
CHUNHUI JI
, 
XIAOZHEN ZHU
XIAOZHEN ZHU
, 
QINGLING MENG
QINGLING MENG
, 
XIANZHU XIA
XIANZHU XIA
, 
XINGXING ZHANG
XINGXING ZHANG
, 
XUEPENG CAI
XUEPENG CAI
, 
KUOJUN CAI
KUOJUN CAI
 e 
QIAO JUN
QIAO JUN
   | 20 dic 2021
Polish Journal of Microbiology's Cover Image
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Article Category: original-paper
Pubblicato online: 20 dic 2021
Pagine: 479 - 487
DOI: https://doi.org/10.33073/pjm-2021-045
Parole chiave
© 2021 Zhongmei Ma et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Molecular characteristics of STM0859 of Salmonella Typhimurium; A) the H-T-H DNA binding and LysR substrate binding domains in STM0859 protein, B) prediction of 3D structure model of STM0859 protein.
Molecular characteristics of STM0859 of Salmonella Typhimurium; A) the H-T-H DNA binding and LysR substrate binding domains in STM0859 protein, B) prediction of 3D structure model of STM0859 protein.

Fig. 2.

Growth curves of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 under different conditions; A-F) growth curves of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 at 37°C, pH 4, 0.3% cholate, 4% NaCl, 1 mM H2O2 and pH 10, respectively.
Growth curves of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 under different conditions; A-F) growth curves of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 at 37°C, pH 4, 0.3% cholate, 4% NaCl, 1 mM H2O2 and pH 10, respectively.

Fig. 3.

Assay of motility of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 in semi-solid medium with different pH; ns – not significant, while ** – p < 0.01.
Assay of motility of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 in semi-solid medium with different pH; ns – not significant, while ** – p < 0.01.

Fig. 4.

Determination of formatting capability of biofilm of SL1344, ΔSTM0859 and ΔSTM0859/STM0859; A) formation of biofilm by SL1344, ΔSTM0859 and ΔSTM0859/STM0859 at 30 h, and 48 h, respectively; B) biofilm of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 biofilm determined by OD595 nm at 30 h, and 48 h, respectively; ns – not significant, * – p < 0.05, while ** – p < 0.01.
Determination of formatting capability of biofilm of SL1344, ΔSTM0859 and ΔSTM0859/STM0859; A) formation of biofilm by SL1344, ΔSTM0859 and ΔSTM0859/STM0859 at 30 h, and 48 h, respectively; B) biofilm of SL1344, ΔSTM0859 and ΔSTM0859/STM0859 biofilm determined by OD595 nm at 30 h, and 48 h, respectively; ns – not significant, * – p < 0.05, while ** – p < 0.01.

Fig. 5.

Determination of the relative transcriptional levels of rpoS, rcsB and fur gene; ns – indicates not significant, * – p < 0.05, while ** – p < 0.01.
Determination of the relative transcriptional levels of rpoS, rcsB and fur gene; ns – indicates not significant, * – p < 0.05, while ** – p < 0.01.

Fig. 6.

Analysis of the interaction between STM0859 and the promoter sequence of rcsB gene using EMSA; A) the relevant features and localization in genomic DNA of ST, B) the promoter sequence of rcsB gene used in this study, C) interaction between STM0859 and the promoter of rcsB gene, D) interaction between BSA and the promoter of rcsB gene (negative control).
Analysis of the interaction between STM0859 and the promoter sequence of rcsB gene using EMSA; A) the relevant features and localization in genomic DNA of ST, B) the promoter sequence of rcsB gene used in this study, C) interaction between STM0859 and the promoter of rcsB gene, D) interaction between BSA and the promoter of rcsB gene (negative control).

The primers used in the study.

Primer namePrimer sequence (5’->3’)Target gene/DNA fragmentProduct size (bp)
R1TCTATGACAAGGATGAAGCCpKD461,046
R2GTATTTCCCGGCCTTTCTGT
R3ACTACGCCTGAAGGTGGCAACGGACCAAATATACAGGGATGTAACGCTATTAGAGCCTCTCAAAGCAATpKD31,198
R4TCTATAAAAAAGCAGCACACTGTATTATACGTTAATTATGAGCCACAACGGTCTTGAGCGATTGTGTAGG
R5ACAACTCAAAGGAAAAGGACpCP201,202
R6CACCAAAAACTCGTAAAAGC
R7GACCAAATTATACAGGGATGTST-ΔSTM0859cat/ST-SL1344220/1,146
R8ATACGTTAATTATGAGCCACA
R9GGAATTCATGCACTTTGATATAAAAGATTTASTM0859905
R10CCTCGAGTCACTCAGTCTGAGCTGTGAG
rcsB FAAAATGCCGGATGGCGGrcsB promoter102
rcsB RTCGCTTTTATGTTACCCAGCC
16s RNA FGCGGTAATACGGAGGGTG16s RNA167
16s RNA RCACCGCTACACCTGGAAT
rcsB FAGCGTCATTTTCCGAGCrcs B193
rcsB RTCCAGCAGACGAGAAACG
rpoS FGGTGGATTCGCCAGACAArpoS132
rpoS RCGTGGTCCAGTTTATGCG
Fur FTTTTCCGCAATCAAGGCFur158
Fur RTGGGTGAAGAAATCGGTC
eISSN:
2544-4646
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Inglese
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