Role of the methylcitrate cycle in propionate metabolism and detoxification in Mycobacterium smegmatis.
Catabolism of odd-chain-length fatty acids yields acetyl-CoA and propionyl-CoA. A common pathway of propionyl-CoA metabolism in micro-organisms is the methylcitrate cycle, which includes the dedicated enzymes methylcitrate synthase (MCS), methylcitrate dehydratase (MCD) and methylisocitrate lyase (MCL). The methylcitrate cycle is essential for propionate metabolism in Mycobacterium tuberculosis. Unusually, M. tuberculosis lacks an MCL orthologue and this activity is provided instead by two isoforms of the glyoxylate cycle enzyme isocitrate lyase (ICL1 and ICL2). These bifunctional (ICL/MCL) enzymes are jointly required for propionate metabolism and for growth and survival in mice. In contrast, the non-pathogenic species Mycobacterium smegmatis encodes a canonical MCL enzyme in addition to ICL1 and ICL2. The M. smegmatis gene encoding MCL (prpB) is clustered with genes encoding MCS (prpC) and MCD (prpD). Here we show that deletion of the M. smegmatis prpDBC locus reduced but did not eliminate MCL activity in cell-free extracts. The residual MCL activity was abolished by deletion of icl1 and icl2 in the DeltaprpDBC background, suggesting that these genes encode bifunctional ICL/MCL enzymes. A DeltaprpB Deltaicl1 Deltaicl2 mutant was unable to grow on propionate or mixtures of propionate and glucose. We hypothesize that incomplete propionyl-CoA metabolism might cause toxic metabolites to accumulate. Consistent with this idea, deletion of prpC and prpD in the DeltaprpB Deltaicl1 Deltaicl2 background paradoxically restored growth on propionate-containing media. These observations suggest that the marked attenuation of ICL1/ICL2-deficient M. tuberculosis in mice could be due to the accumulation of toxic propionyl-CoA metabolites, rather than inability to utilize fatty acids per se.
Laboratory of Infection Biology, The Rockefeller University, New York, NY 10021, USA.
Anna M
AM
Upton
Laboratory of Infection Biology, The Rockefeller University, New York, NY 10021, USA.
John D
JD
McKinney
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D019755:Carbon-Carbon Lyases / Q000235:genetics / Q000378:metabolism; D019343:Citric Acid / Q000378:metabolism*; D019295:Computational Biology; D003470:Culture Media; D017353:Gene Deletion; D005816:Genetic Complementation Test; D007522:Isocitrate Lyase / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D020102:Mycobacterium smegmatis / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism / Q000472:pathogenicity*; D009169:Mycobacterium tuberculosis / Q000235:genetics; D011422:Propionates / Q000378:metabolism*; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D003470:Culture Media; D007527:Isoenzymes; D011422:Propionates; D019343:Citric Acid; D019755:Carbon-Carbon Lyases; D007522:Isocitrate Lyase; C417277:methylisocitrate lyase