A KU55933 purchase hypothetical protein (MAP0860c) upregulated in the presence of iron in the cattle strain of MAP has been described as a part of MAP-specific large sequence polymorphism (LSP4) [22]. Table 3 Transcript and protein expression in cattle MAP under iron-replete (HI) conditions MAP ORF ID Predicted function aFold change Protein Transcript Metabolism MAP0150c FadE25_2 (acyl-coA dehydrogenase) 1.72 ± 0.1 1.88 ± 0.2 MAP0789 acetyl-CoA acetyltransferase 1.73 ±
0.3 1.56 ± 0.1 MAP1846c ATP phosphoribosyltransferase 1.69 ± 0.2 3.68 ± 0.3 MAP2332c Fas (fatty acid synthase) 1.61 ± 0.5 2.28 ± 0.4 MAP3404 AccA3 (acetyl-/propionyl-coenzyme A) 1.45 ± 0.1 2.18 ± 0.2 MAP3698c succinate dehydrogenase 1.89 ± 0.3 4.57 ± 0.5 Cellular processes MAP1339 iron regulated conserved protein 1.62 ��-Nicotinamide ± 0.2 0.78 ± 0.3 MAP1653 thiol peroxidase 1.79 ± 0.5 2.29 ± 0.2 Information storage and processing MAP2907c translation initiation factor IF-2 1.57 ± 0.2 1.89 ± 0.2 MAP2945c ribosome releasing factor 1.66 ± 0.3 2.11 ± 0.5 MAP4113 50S ribosomal
protein L1 1.61 ± 0.1 1.57 ± 0.2 MAP4125 rplJ 50S ribosomal protein L10 1.52 ± 0.1 1.66 ± 0.5 MAP4142 fusA elongation factor G 2.13 ± 0.4 3.05 ± 0.3 MAP4160 rpsJ 30S ribosomal protein S10 1.68 ± 0.3 2.87 ± 0.4 MAP4181 rpsH 30S ribosomal protein S8 1.79 ± 0.5 2.42 ± 0.1 MAP4233 rpoA DNA-directed RNA polymerase 1.56 ± 0.1 1.65 ± 0.4 Poorly characterized pathways MAP0216 FbpA antigen 85-A 1.87 ± 0.2 2.16 ± 0.3 MAP1122 mycobacterial PF-01367338 cell line integration host factor 1.73 ± 0.3 2.00 ± 0.5 a MAP oligoarray was used to measure gene expression whereas iTRAQ was used to quantitate protein expression in the cultures of cattle MAP strain grown in iron-replete (HI) or iron-limiting (LI) medium. Fold change for each target
was calculated and represented as a log2 ratio of HI/LI. Shown are the MAP genes that demonstrated the presence of 1.5 times or more of transcripts and proteins in HI compared to LI. Genes are annotated based on the motif searches in KEGG database. In contrast, we did not document any upregulation (at a log2 fold change of Ureohydrolase 1.5) in the S MAP under iron-replete conditions. The directionality of transcripts as identified by microarrays under iron-replete conditions by S MAP strain was confirmed by real time RT-PCR (Additional file 1, Table S4). Proteome The following criteria were used for protein identification in each treatment – (1) peptides identified by mass spectrometry were searched against the non-redundant (nr) protein database deposited in NCBI); and (2) MAP specific peptides reported with >95% confidence were used to quantify the relative abundance (iron-replete v/s iron-limitation) of each protein. A peptide with no hits on the MAP genome but with identities with other mycobacterial proteins was considered as unannotated MAP protein.