Gene Rv2215
in Mycobacterium tuberculosis H37Rv
General annotation
| Type | CDS |
| Function | Involved in tricarboxylic acid cycle; converts pyruvate to acetyl-CoA and CO2. Also involved in antioxidant defense; LPDC|Rv0462, DLAT|Rv2215, AHPD|Rv2429, and AHPC|Rv2428 constitute an NADH-dependent peroxidase and peroxynitrite reductase that provides protection against oxidative stress. |
| Product | DlaT, dihydrolipoamide acyltransferase, E2 component of pyruvate dehydrogenase |
| Comments | Rv2215, (MTCY190.26), len: 553 aa. DlaT, dihydrolipoamide acyltransferase, E2 component of pyruvate dehydrogenase, proven biochemically (see Tian et al. 2005), similar to e.g. SW:O PD2_ACHLA P35489 dihydrolipoamide acetyltransferase component (E2) of pyruvate dehydrogenase complex (35.3% identity in 552 aa overlap); contains PS00189 2-oxo acid dehydrogenases acyltransferase component lipoyl binding site. Rhodanine compounds inhibit DlaT|Rv2215 and can kill non-replicating mycobacteria in mouse bone marrow-derived macrophages (See Bryk et al., 2008). LpdC|Rv0462 co-immunoprecipitates with DlaT|Rv2215 (in lpdC|Rv0462 mutant) and with BkdC|Rv2495c (in dlaT|Rv2215 mutant) (See Venugopal et al., 2011). |
| Functional category | Intermediary metabolism and respiration |
| Proteomics | Identified by proteomics (See Rosenkrands et al., 2000). Identified in the membrane fraction of M. tuberculosis H37Rv using 1D-SDS-PAGE and uLC-MS/MS (See Gu et al., 2003). Identified in the cytosol and cell membrane fraction of M. tuberculosis H37Rv using 2DLC/MS (See Mawuenyega et al., 2005). Identified by mass spectrometry in Triton X-114 extracts of M. tuberculosis H37Rv (See Malen et al., 2010). Identified by mass spectrometry in the membrane protein fraction and whole cell lysates of M. tuberculosis H37Rv but not the culture filtrate (See de Souza et al., 2011). Translational start site supported by proteomics data (See de Souza et al., 2011) (See Kelkar et al., 2011). |
| Mutant | Essential gene for in vitro growth of H37Rv in a MtbYM rich medium, by Himar1 transposon mutagenesis (see Minato et al. 2019). Non-essential gene for in vitro growth of H37Rv, by analysis of saturated Himar1 transposon libraries (see DeJesus et al. 2017). Essential gene by Himar1 transposon mutagenesis in H37Rv strain (see Sassetti et al., 2003). Essential gene for in vitro growth of H37Rv, by Himar1 transposon mutagenesis (See Griffin et al., 2011). M. tuberculosis H37Rv dlaT|Rv2215 mutant (gene replacement) has severe growth defect in vitro, has increased sensitivity to nitrosative stress but not heat or INH; mutant shows reduced survival in bone marrow-derived macrophages from C57BL/6 and iNOS -/- mice; growth of mutant is impaired in C57BL/6 mouse lungs, spleen and liver (See Shi and Ehrt, 2006). Hartley guinea pigs were infected with M. tuberculosis H37Rv, M. tuberculosis H37Rv dlaT|Rv2215 mutant, or complemented mutant - wild-type and complemented strains caused severe tuberculosis while there was no detectable disease with the mutant; growth of the mutant in lungs and spleen is impaired (See Bryk et al., 2008). In 7H9 medium, M. tuberculosis H37Rv dlaT|Rv2215 mutant survives into stationary phase while lpdC|Rv0462 mutant dies, but growth curves are similar to wild-type if dextrose and glycerol are removed; lpdC|Rv0462 and dlaT|Rv2215 mutants have elevated levels of pyruvate and derived amino acids which return to near-wild-type levels by 28 days in dlaT|Rv2215 mutant but not lpdC|Rv0462 mutant; dlaT|Rv2215 mutant shows increased growth in 7H9 with leucine or isoleucine while lpdC|Rv0462 mutant and bkdC|Rv2495c-dlaT|Rv2215 double mutant show little or no growth; bkdA|Rv2497c (qRT-PCR) and bkdC|Rv2495c (qRT-PCR, Western) are upregulated in dlaT|Rv2215 mutant; lysates from dlaT|Rv2215 mutant have branched-chain keto acid dehydrogenase activity while those of wild-type and lpdC|Rv0462 mutant do not; growth of bkdC|Rv2495c-dlaT|Rv2215 double mutant in C57BL/6 lungs is strongly attenuated (See Venugopal et al., 2011). Check for mutants available at TARGET website |
Coordinates
| Type | Start | End | Orientation |
|---|---|---|---|
| CDS | 2481965 | 2483626 | + |
Genomic sequence
Feature type
Upstream flanking region (bp)
Downstream flanking region (bp)
Update
Protein sequence
>Mycobacterium tuberculosis H37Rv|Rv2215|dlaT
MAFSVQMPALGESVTEGTVTRWLKQEGDTVELDEPLVEVSTDKVDTEIPSPAAGVLTKIIAQEDDTVEVGGELAVIGDAKDAGEAAAPAPEKVPAAQPESKPAPEPPPVQPTSGAPAGGDAKPVLMPELGESVTEGTVIRWLKKIGDSVQVDEPLVEVSTDKVDTEIPSPVAGVLVSISADEDATVPVGGELARIGVAADIGAAPAPKPAPKPVPEPAPTPKAEPAPSPPAAQPAGAAEGAPYVTPLVRKLASENNIDLAGVTGTGVGGRIRKQDVLAAAEQKKRAKAPAPAAQAAAAPAPKAPPAPAPALAHLRGTTQKASRIRQITANKTRESLQATAQLTQTHEVDMTKIVGLRARAKAAFAEREGVNLTFLPFFAKAVIDALKIHPNINASYNEDTKEITYYDAEHLGFAVDTEQGLLSPVIHDAGDLSLAGLARAIADIAARARSGNLKPDELSGGTFTITNIGSQGALFDTPILVPPQAAMLGTGAIVKRPRVVVDASGNESIGVRSVCYLPLTYDHRLIDGADAGRFLTTIKHRLEEGAFEADLGL
Bibliography
- Rosenkrands I et al. [2000]. Towards the proteome of Mycobacterium tuberculosis. Proteomics
- Bryk R et al. [2002]. Metabolic enzymes of mycobacteria linked to antioxidant defense by a thioredoxin-like protein. Function
- Sassetti CM et al. [2003]. Genes required for mycobacterial growth defined by high density mutagenesis. Mutant
- Gu S et al. [2003]. Comprehensive proteomic profiling of the membrane constituents of a Mycobacterium tuberculosis strain. Proteomics
- Tian J et al. [2005]. Variant tricarboxylic acid cycle in Mycobacterium tuberculosis: identification of alpha-ketoglutarate decarboxylase. Biochemistry
- Tian J, Bryk R, Shi S, Erdjument-Bromage H, Tempst P and Nathan C [2005]. Mycobacterium tuberculosis appears to lack alpha-ketoglutarate dehydrogenase and encodes pyruvate dehydrogenase in widely separated genes. Biochemistry
- Mawuenyega KG et al. [2005]. Mycobacterium tuberculosis functional network analysis by global subcellular protein profiling. Proteomics
- Shi S et al. [2006]. Dihydrolipoamide acyltransferase is critical for Mycobacterium tuberculosis pathogenesis. Mutant
- Bryk R et al. [2008]. Selective killing of nonreplicating mycobacteria. Biochemistry Mutant
- Målen H et al. [2010]. Definition of novel cell envelope associated proteins in Triton X-114 extracts of Mycobacterium tuberculosis H37Rv. Proteomics
- Kelkar DS et al. [2011]. Proteogenomic analysis of Mycobacterium tuberculosis by high resolution mass spectrometry. Proteomics Sequence
- Venugopal A et al. [2011]. Virulence of Mycobacterium tuberculosis depends on lipoamide dehydrogenase, a member of three multienzyme complexes. Biochemistry Mutant
- de Souza GA et al. [2011]. Bacterial proteins with cleaved or uncleaved signal peptides of the general secretory pathway. Proteomics
- Griffin JE et al. [2011]. High-resolution phenotypic profiling defines genes essential for mycobacterial growth and cholesterol catabolism. Mutant
- de Souza GA et al. [2011]. Proteogenomic analysis of polymorphisms and gene annotation divergences in prokaryotes using a clustered mass spectrometry-friendly database. Proteomics Sequence
- DeJesus MA et al. [2017]. Comprehensive Essentiality Analysis of the Mycobacterium tuberculosis Genome via Saturating Transposon Mutagenesis. Mutant
- Minato Y et al. [2019]. Genomewide Assessment of Mycobacterium tuberculosis Conditionally Essential Metabolic Pathways. Mutant
