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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.