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Supplemental Tables & Figures
Table S1. Gene expression of MAC104 microaggregates by microarray analysis in
comparison to planktonic bacteria.
Locus Taga
MAV_3013
MAV_5199
MAV_0345
MAV_2086
MAV_4089
MAV_2085
MAV_4081
MAV_4088
MAV_4082
MAV_4086
MAV_0291
MAV_4074
Accession no.
YP_882199
YP_884315
YP_879632
YP_881301
-YP_881300
YP_883231
YP_883238
YP_883232
YP_883236
YP_879580
YP_883225
MAV_4087
MAV_1799
MAV_4070
MAV_0831
MAV_4085
MAV_1331
YP_883237
YP_881023
YP_883221
YP_880104
YP_883235
YP_880576
MAV_4092
MAV_1332
MAV_4086
MAV_2107
MAV_3093
YP_883241
YP_880577
YP_883236
YP_881318
YP_882277
MAV_3159
MAV_2087
MAV_3092
MAV_4090
YP_882343
YP_881302
YP_882276
YP_883239
MAV_4091
YP_883240
Putative Identification
hypothetical protein
hypothetical protein
methylisocitrate lyase
ornithine--oxo-acid transaminase
pseudogene
amidinotransferase superfamily protein
transferase
phosphoheptose isomerase
transferase
transferase
hypothetical protein
GlcNAc-PI de-N-acetylase family
protein
transferase
hypothetical protein
hypothetical protein
hypothetical protein
transferase
1-pyrroline-5-carboxylate
dehydrogenase
UDP-glucose 6-dehydrogenase
proline dehydrogenase
transferase
hypothetical protein
glucose-methanol-choline
oxidoreductase
hypothetical protein
amino acid transporter
hypothetical protein
oxidoreductase, short chain
dehydrogenase/reductase family
protein
transferase
Fold Change
6.47
5.97
5.56
5.07
4.91
4.78
4.75
4.72
4.59
4.57
4.48
4.25
4.17
4.12
4.09
4.06
3.91
3.91
3.88
3.84
3.83
3.82
3.76
3.70
3.68
3.65
3.62
3.59
MAV_4063
YP_883215
MAV_0439
MAV_3734
MAV_0787
MAV_3735
MAV_3201
MAV_2581
MAV_3091
MAV_0675
MAV_4064
-YP_882908
YP_880060
YP_882909
-YP_881772.1
YP_882275
YP_879954
YP_883216
MAV_3202
MAV_4100
MAV_4093
MAV_1854
MAV_4069
MAV_2749
YP_882384
YP_883249
YP_883242
YP_881076
YP_883220
YP_881938
MAV_3882
MAV_3511
MAV_2386
MAV_1736
MAV_4073
MAV_3281
MAV_2381
MAV_0789
MAV_3930
MAV_3656
MAV_4536
MAV_2387
MAV_2277
MAV_3617
MAV_3310
MAV_2707
MAV_0427
MAV_3347
YP_883044
YP_882693
YP_881586
YP_880963
YP_883224
YP_882463
YP_881582
YP_880062
YP_883092
YP_882833
YP_883667
YP_881587
YP_881481
YP_882793
YP_882492
-YP_879711
YP_882530
MAV_2751
MAV_2471
YP_881940
YP_881663
transcriptional regulator, AraC family
protein
pseudogene
hypothetical protein
gp50 protein
transcriptional regulator GlnR
pseudogene
hypothetical protein
hypothetical protein
hypothetical protein
glycosyl transferase, group 1 family
protein
transmembrane protein
glycogen debranching enzyme GlgX
pyruvate decarboxylase
transposase Integrase core domain
catalase HPII
FAD dependent oxidoreductase
domain-containing protein
hypothetical protein
dihydrodiol dehydrogenase
virulence factor Mce family protein
isochorismatase family protein
dehydrogenase
hypothetical protein
hypothetical protein
hypothetical protein
hypothetical protein
hypothetical protein
hypothetical protein
virulence factor Mce family protein
hypothetical protein
hypothetical protein
27 kDa lipoprotein antigen
pseudogene
methylase, putative
transposaseIclR helix-turn-helix
domain
ErfK/YbiS/YcfS/YnhG family protein
DoxX subfamily protein, putative
3.51
3.50
3.49
3.43
3.42
3.33
3.31
3.29
3.24
3.20
3.20
3.18
3.16
3.13
3.12
3.11
3.01
2.96
2.95
2.95
2.94
2.90
2.90
2.89
2.88
2.87
2.87
2.79
2.77
2.70
2.68
2.68
2.67
2.66
2.65
2.65
MAV_3982
YP_883138
MAV_2383
MAV_2112
MAV_3094
MAV_3739
MAV_1565
MAV_2384
MAV_1978
MAV_2390
MAV_4076
MAV_2072
MAV_2780
MAV_1856
MAV_2380
MAV_0752
MAV_3737
MAV_0438
MAV_4078
MAV_0344
MAV_4067
MAV_1660
MAV_3095
MAV_0244
MAV_3505
YP_881583
-YP_882278
-YP_880799
YP_881584
YP_881197
YP_881590
YP_883226
YP_881288
YP_881968
YP_881078
YP_881581
YP_880027
YP_882911
YP_879722
YP_883229
YP_879631
YP_883219
YP_880893
YP_882279
YP_879535
YP_882687
MAV_2429
MAV_2760
MAV_4072
MAV_2372
MAV_2002
MAV_0426
YP_881625
YP_881949
YP_883223
YP_881574
YP_881221
YP_879710
MAV_5176
MAV_0957
MAV_2388
MAV_3663
MAV_5222
MAV_0440
MAV_4256
YP_884293
YP_880217
YP_881588
YP_882840
-YP_879723
YP_883398
hemerythrin HHE cation binding
domain-containing protein
mce related protein
psedogene
glycosyl hydrolase, family protein 15
pseudogene
hypothetical protein
mce related protein
hypothetical protein
hypothetical protein
endonuclease/exonuclease/phosphotase
hypothetical protein
hypothetical protein
transposase
transposase
transposase
hypothetical protein
hypothetical protein
hypothetical protein
methylcitrate synthase
stas domain-containing protein
hypothetical protein
short chain dehydrogenase
hypothetical protein
protocatechuate 3,4-dioxygenase,
alpha subunit
PPE family protein
hypothetical protein
hypothetical protein
hypothetical protein
transposase
intracellular protease, PfpI family
protein
hypothetical protein
hypothetical protein
mce related protein
hypothetical protein
pseudogene
restriction endonuclease family protein
RNA polymerase sigma factor SigF
2.62
2.62
2.61
2.58
2.57
2.57
2.56
2.54
2.54
2.54
2.53
2.52
2.51
2.49
2.47
2.47
2.45
2.45
2.44
2.43
2.43
2.41
2.39
2.39
2.37
2.36
2.35
2.33
2.30
2.30
2.29
2.28
2.24
2.23
2.22
2.21
2.21
MAV_3011
YP_882197
MAV_2702
MAV_3978
MAV_2701
MAV_0122
MAV_3892
MAV_4982
YP_881893
YP_883134
YP_881892
YP_879418
YP_883054
YP_884103
MAV_4077
MAV_0302
MAV_0346
MAV_2077
MAV_4095
YP_883228
YP_879591
YP_879633
YP_881293
YP_883244
MAV_1412
MAV_4141
MAV_3407
MAV_1735
MAV_3940
MAV_1340
MAV_3103
MAV_2170
MAV_1624
MAV_2706
YP_880653
YP_883289
YP_882589
YP_880962
MAV_4101
YP_883250
MAV_3096
YP_882280
MAV_2704
YP_881895
MAV_1040
MAV_3881
MAV_2385
MAV_2587
MAV_0806
MAV_2955
MAV_3891
MAV_0811
YP_880296
YP_883043
YP_881585
YP_881778
YP_880079
YP_882141
YP_883053
YP_880084
YP_880584
YP_882287
YP_881380
YP_880857
YP_881897
alpha/beta hydrolase domaincontaining protein
acetyl-CoA acetyltransferase
hypothetical protein
putative acyl-CoA dehydrogenase
hypothetical protein
hypothetical protein
short-chain dehydrogenase/reductase
SDR
hypothetical protein
hypothetical protein
2-methylcitrate dehydratase 2
hypothetical protein
hemerythrin HHE cation binding
domain-containing protein
hypothetical protein
ABC1 family protein
hypothetical protein
NAD-dependent deacetylase 1
psedogene
aldehyde dehydrogenase
ANTAR domain-containing protein
acyl carrier protein
hypothetical protein
hydrolase, alpha/beta fold family
protein, putative
S-(hydroxymethyl)glutathione
dehydrogenase
mandelate racemase/muconate
lactonizing enzyme
L-carnitine dehydratase/bile acidinducible protein F
hypothetical protein
hypothetical protein
virulence factor Mce family protein
AMP-dependent synthetase and ligase
hypothetical protein
hypothetical protein
methylesterase
hypothetical protein
2.20
2.20
2.19
2.19
2.18
2.18
2.17
2.16
2.16
2.16
2.16
2.15
2.13
2.13
2.12
2.11
2.10
2.09
2.09
2.08
2.07
2.07
2.07
2.07
2.07
2.04
2.03
2.03
2.02
2.02
2.02
2.02
2.01
MAV_0788
MAV_3929
MAV_1513
a
YP_880061
YP_883091
YP_880753
hypothetical protein
hypothetical protein
hypothetical protein
2.01
2.01
2.00
Locus tags were obtained from the MAC104 genome as deposited in the National
Center for Biotechnology Information (NCBI) database.
Figure S1. Differentially expressed genes during MAH microaggregate formation
categorized based on the known or predicted functions of the encoded proteins. Only
genes with a 2-fold change or greater were included.
Table S2. Confirmation of microarray gene expression by real-time PCR.
Fold difference (microaggregate vs planktonic
cells)
Locus tag
Putative
identification
Microarray
RT-PCR
MAV_3013
MAV_1799
MAV_5199
MAV_0831
Hypothetical Protein
Hypothetical Protein
Hypothetical Protein
Hypothetical Protein
6.47
4.12
5.97
4.06
1.8 ± 0.12
3.0 ± 1.3
1.5 ± 0.3
2.4 ± 0.9
a
a
Locus tags were obtained from the MAC104 genome as deposited in the National
Center for Biotechnology Information (NCBI) database.
Figure S2. Multiple sequence alignment of MBP-1 protein with human, rat and
mouse ADIP protein produced by UniProt KB. Square box indicates the amino acids
that were deleted to create the MBP-1 dominant-negative construct, Msmeg-3013Δ45-63.
[*] denotes identical residues in all sequences. [:] denotes conserved substitutions [.]
denotes semi-conserved substitutions.
Figure S3. Confirmation of anti-MBP-1 immune serum. (1) BL21 E. coli lysate, (2)
non-specific protein (Rv3345) (3) MBP-1 overexpressed in BL21 E. coli lysate