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Table 2 Summary of survival characteristics of novel attenuated mutants

From: Mariner mutagenesis of Brucella melitensis reveals genes with previously uncharacterized roles in virulence and survival

Chr BME GENE FUNCTION Gene S v. R a In vitro Survival Ratio M Survival Ratio c Mouse Survival Ratio c,d
Intracellular trafficking, secretion, and vesicular transport
I 1894 Gramicidin S biosynthesis grsT S -0.4 1.5 0.9
II 0029–30 VirB5-B6 virB5/6 S    
II 0034 Type IV secretion (channel protein) virB10 S   2.5*  
II 0035 Type IV secretion (ATPase) virB11 S   2.3***  
Nucleotide transport and metabolism
I 0233 Phosphoribosylaminoimidazolecarboxamide formyltransfease purH S 0.4 0.9  
I 0295 Phosphoribosylaminoimidazole carboxylase ATPase subunit purK S 0.3 0.9  
I 1123 Phosphoribosylformylglycinamidine (FGAM) synthase purS S 0.9 2.9  
I 1124 Phosphoribosylformylglycinamidine (FGAM) synthase purQ S 1.9 2.2  
I 1204 Exopolyphosphatase (Ppx/GppA) ppx S 0.4 1.2  
Signal transduction mechanisms
I 0066 Response regulator (two component) ompR S 0.6 1.3 1.9*
I 1327 Glutamate-ammonia-ligase adenylyltransferase (signal transduction) glnE S -0.1 1.2 1.4
I 1807 Universal stress protein uspA S   0.4 0.1
Transcription
I 0304/5 Hypothetical thioesterase-transcriptional regulator deoR S -0.5 1.9  
I 0371 Regulatory factor (rpoE) virF S -0.1 1.1 0.3
I 0508 Transcription elongation factor greA S 1.0 0.7 0.6
I 0513 Transcription factor lysR S   0.9  
I 0513/4 Transcription factor-hypothetical protein lysR/hyp S 0.2 0.7  
I 0731/2 Cold shock protein-hypothetical cytosolic protein csp/hyp S 0.2 1.1 0.9
I 0808 Transcriptional regulator (soxR) merR S 1.0 0.8  
I 1178 Transcriptional regulator (soxR) merR S 0.0   
I 1297/8 RNA polymerase omega subunit-hypothetical protein rpoZ/hyp S 0.2 1.3  
I 1364 Transcriptional regulatory protein mucR S 0.3 2.0*** 3.0**
I 1647/8 Hyp-RegM (ATP-binding) hyp/regM S   1.0 -0.2
II 1116/7 Transcriptional activator LuxR-TetR families luxR/tetR S 0.1 1.6** 1.1
Cell wall/membrane/envelope biogenesis
I 0498/9 Cold shock protein/soluble lytic transglycosylase cspA/slt R 0.0 1.7*** 2.9
I 1302 Soluble lytic murein transglycosylase mltE R 0.6 0.7 0.4
I 1415 O-antigen export system permease protein rfbD R 0.1 1.3  
I 1416 O-antigen export system ATP-binding protein rfbE R 0.3 1.9**  
I 1427 UDP-N-acetylglucosamine 4,6-dehydratase capD R -0.3 1.4  
II 0380 Acriflavin resistance protein A (membrane biogenesis) acrA(hlyD) S   3.4  
II 0472 Membrane fusion protein (membrane biogenesis) mtrC(hlyD) S 0.9 0.3  
Energy production and conversion
I 0972 Glutathione reductase gor S 0.3 0.5  
I 1749 FAD-dependent glycerol phosphate dehydrogenase glpD S 0.2 0.7  
Amino acid transport and metabolism
II 0285 Dipeptide transport system (inner membrane permease) dppB S   0.4 0.7
Replication, recombination and repair
I 0334 AAA ATPase/DNA helicase/DNA damage checkpoint ruvB S 1.3 0.9  
I 1307 Integrase/recombinase (phage) xerC (int) S 0.1 1.1  
I 2023 ATP-dependent nuclease subunit A uvrD/rep S   0.9  
II 0260 GTP-binding protein lepA S 0.9 0.1  
I 1296 GTP-binding protein lepA S   1.0  
Translation, ribosome structure and biogenesis
I 0983 Ribosomal large subunit pseudouridine synthase C rluA S 0.5 1.1 1.0
I 1057 Ribonuclease E/Zn metalloprotease cafA S 0.5 1.1 1.6
I 1775–6 Rnase PH-HrcA (heat shock protein repressor) rph/hrcA S -0.2 1.0 0.6
Function unknown
I 0186 Hypothetical cytosolic protein dut S 0.6 0.4  
I 0193 Hypothetical protein hyp S 0.4 2.4 1.5
I 0490 Hypothetical cytosolic protein hycp S 0.6 0.6  
I 0540 Hypothetical protein hyp S 0.0 0.6  
I 0603 Hypothetical protein hyp S -0.1 1.1  
I 0732 Hypothetical cytosolic protein hycp S   1.4  
II 0045–6 Hypothetical protein-Hypothetical protein hyp/hyp S    
Posttranslational modification, protein turnover and chaperones
I 0816 ATP-dependent clp protease ATP-binding subunit clpA S 1.0 0.8 1.5
II 0932 Glutaredoxin nrdH S   1.1 -0.3
Defense mechanisms
I 0926 Multidrug resistance locus emrA    2.2  
General function prediction
I 1143 Metal-dependent hydrolase (β-lactamase) mbl S -0.1 0.9  
I 1282 Glycine cleavage T protein (aminomethyltransferase) gcvT S 1.2 0.4  
I 1487 Colicin V production protein colV S 1.2 0.5 0.8
I 1499 Pirin (cupin) pirA S -0.1 1.3 1.7
I 1867 Florfenicol resistance protein nifB/elp3 S 0.4 0.5  
Carbohydrate transport and metabolism
I 2031 Phosphocarrier protein HPr ptsH S -0.3 0.9 0.4
II 1095 L-fuculose phosphate aldolase sbgE S 0.7   2.2*
Lipid transport and metabolism
Coenzyme transport and metabolism
I 0657 Outer membrane receptor (TonB-dependent transport) btuB S 0.2 0.6  
  1. a S v. R denotes smooth versus rough colony morphology
  2. b In vitro growth: 1:1 mixtures of 16 M and select mutants used to inoculate culture to evaluate growth defects of the mutant. The output ratios are corrected for variance from the input ratios (approximately 1.0) and presented as a log10 value, also refrerred to as Δlog10. Macrophage survival is presented as a log10 of the ratio of recovery 16 M to mutant which may also be presented as Δlog10.
  3. c Statistical analysis of macrophage and mouse splenic clearance revealed significant differences among means using 1-way ANOVA, Dunn's multiple comparisons using a non-attenuated mutant revealed statistically valid differences with p-values of 0.05 (*), 0.01 (**) and 0.001 (***).
  4. d Mouse splenic clearance: 1:1 mixtures of 16 M and select mutants were used to inoculate mice i.p. to evaluate degree of in vivo attenuation of survival at one week.
  5. / Indicates intergenic insertion between two genes.