CopA Protects Streptococcus suis against Copper Toxicity
Abstract
:1. Introduction
2. Results
2.1. S. suis CopA Is a Homologue of the Copper Efflux System
2.2. S. suis Up-regulates copA Expression in Response to Copper
2.3. CopA Is Required for Copper Resistance in S. suis
2.4. copA Deletion Leads to Increased Intracellular Accumulation of Copper
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains, Plasmids, and Growth Conditions
4.2. Bioinformatic Analysis
4.3. copA Expression Analysis
4.4. Construction of the Complementation Strain
4.5. Growth Curve Analyses
4.6. Spot Dilution Assays
4.7. Intracellular Copper Content Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
WT | wild-type |
ICP-OES | inductively coupled plasma-optical emission spectroscopy |
TSBS | Tryptic Soy Broth supplemented with 10% newborn bovine serum |
TSAS | Tryptic Soy Agar supplemented with 10% newborn bovine serum |
PBS | phosphate buffered saline |
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S. suis Strains | Locus Tag | Gene Sequence Identity (%) 1 |
---|---|---|
LSM102 | A9494_06425 | 100 |
SC19 | B9H01_06680 | 100 |
SS2-1 | BVD85_06510 | 100 |
ZY05719 | ZY05719_06610 | 100 |
A7 | SSUA7_1228 | 100 |
P1/7 | SSU1214 | 100 |
BM407 | SSUBM407_0575 | 100 |
SC84 | SSUSC84_1247 | 100 |
S735 | - | 99 |
GZ1 | SSGZ1_1230 | 99 |
SS12 | SSU12_1279 | 99 |
05ZYH33 | SSU05_1385 | 99 |
98HAH33 | SSU98_1400 | 99 |
SH0104 | - | 97 |
HA0609 | CR542_03955 | 97 |
90-1330 | AN924_03380 | 97 |
NSUI060 | APQ97_02765 | 97 |
NSUI002 | AA105_03890 | 97 |
05HAS68 | HAS68_0686 | 97 |
YB51 | YB51_2960 | 97 |
D9 | SSUD9_0599 | 97 |
ST3 | SSUST3_0597 | 97 |
CS100322 | CR541_06915 | 97 |
T15 | T15_0568 | 97 |
SC070731 | NJAUSS_1288 | 97 |
JS14 | SSUJS14_1360 | 97 |
ST1 | SSUST1_0574 | 96 |
ISU2812 | A7J09_03980 | 96 |
SH1510 | DP111_07130 | 96 |
GZ0565 | BFP66_02780 | 95 |
DN13 | A6M16_02880 | 95 |
6407 | ID09_03115 | 95 |
TL13 | TL13_0615 | 95 |
CZ130302 | CVO91_03355 | 95 |
HN105 | DF184_07440 | 95 |
HN136 | CWM22_09360 | 95 |
SRD478 | A7J08_03040 | 92 |
1081 | BKM67_07590 | 93 |
0061 | BKM66_07040 | 93 |
D12 | SSUD12_0568 | 92 |
HA1003 | DP112_07660 | 92 |
AH681 | CWI26_08525 | 92 |
Strain or Plasmid | Relevant Characteristics | Source or Reference |
---|---|---|
Strains | ||
SC19 | Virulent S. suis 2 strain isolated from the brain of a dead pig | [36] |
ΔcopA | copA deletion mutant of strain SC19 | [23] |
ΔcopA::copA | Complemented strain of ΔcopA | This study |
DH5α | Cloning host for recombinant vector | TransGen |
Plasmids | ||
pSET4s | Thermosensitive suicide vector; SpcR 1 | [37] |
pSET4s::CcopA | pSET4s containing copA and its flanking regions | This study |
Primer | Sequence (5’-3’) 1 | Size (bp) | Target Gene |
---|---|---|---|
QcopA1 | AGAGGATAGGGATGAGCAAGATAACT | 148 | an internal region of copA |
QcopA2 | TTTGTCTGGTCAGCAGCATTTACT | ||
Q16S1 | TAGTCCACGCCGTAAACGATG | 159 | an internal region of 16S rRNA |
Q16S2 | TAAACCACATGCTCCACCGC | ||
L1 | CCCCGTCGACAATGAGGGCCAAAACGTC | 3758 | copA and its flanking regions |
R2 | CGCCGAATTCACCATCGACCAGCACTGAG | ||
In1 | TATCACCGAAAGACCACGAC | 629 | an internal region of copA |
In2 | ATAATGTTTTTGGCGGCAC | ||
Out1 | GAGGACAAAATCAGGGGCT | 2769/378 | a fragment containing copA |
Out2 | AGGGAACAGGCTGAAAACC |
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Zheng, C.; Jia, M.; Lu, T.; Gao, M.; Li, L. CopA Protects Streptococcus suis against Copper Toxicity. Int. J. Mol. Sci. 2019, 20, 2969. https://doi.org/10.3390/ijms20122969
Zheng C, Jia M, Lu T, Gao M, Li L. CopA Protects Streptococcus suis against Copper Toxicity. International Journal of Molecular Sciences. 2019; 20(12):2969. https://doi.org/10.3390/ijms20122969
Chicago/Turabian StyleZheng, Chengkun, Mengdie Jia, Tianyu Lu, Miaomiao Gao, and Lingzhi Li. 2019. "CopA Protects Streptococcus suis against Copper Toxicity" International Journal of Molecular Sciences 20, no. 12: 2969. https://doi.org/10.3390/ijms20122969