Moonlighting Glutamate Formiminotransferases Can Functionally Replace 5-Formyltetrahydrofolate Cycloligase

Jeanguenin, Linda; Lara‐Núñez, Aurora; Pribat, Anne; Magerøy, Melissa H.; Gregory, Jesse F.; Rice, Kelly C.; Crécy‐Lagard, Valérie de; Hanson, Andrew D.

doi:10.1074/jbc.m110.190504

Cited by 26 publications

(30 citation statements)

References 59 publications

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“…S1). Interestingly, the 6S RNA genes are often located downstream of purK (encoding phosphoribosylaminoimidazole carboxylase), which is reminiscent of the situation in many entero-and c-proteobacteria, where the 6S RNA gene is co-transcribed with the ygfA gene, also involved in purine metabolism (Barrick et al, 2005;Jeanguenin et al, 2010).…”

Section: Resultsmentioning

confidence: 99%

6S RNA – an old issue became blue-green

et al. 2012

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Section: Resultsmentioning

confidence: 99%

6S RNA – an old issue became blue-green

et al. 2012

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“…Compared with M. smegmatis, deletion of ygfA in E. coli resulted in a lower reduction of antifolate resistance (Table 1) and only a 6-fold increase in total cellular content of 5-CHO-H 4 PteGlu n (39). The reason behind this species-specific difference is currently unclear.…”

Section: Discussionmentioning

confidence: 82%

“…The reason behind this species-specific difference is currently unclear. A recent study showed that conversion of 5-CHO-H 4 PteGlu n (to free H 4 PteGlu n ) could be catalyzed by the moonlighting activity of glutamate formiminotransferase (FT) in some bacteria, especially those lacking MTHFS activity (39). This enzyme might thus help those bacteria to compensate for the absence of MTHFS by lowering the toxic accumulation of 5-CHO-H 4 PteGlu n species.…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, both E. coli and mycobacteria (Ref. 39 and our BLAST search) do not have genes encoding formiminotransferase homologs; thus, they are unlikely to be able to recycle 5-CHO-H 4 PteGlu n by the moonlighting activity of formiminotransferase. The precise mechanism(s) underlying the role of MTHFS in bacterial antifolate resistance thus remains to be further characterized.…”

Section: Discussionmentioning

confidence: 99%

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Bacterial Conversion of Folinic Acid Is Required for Antifolate Resistance

Ogwang

Nguyen

Sherman

et al. 2011

Journal of Biological Chemistry

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Antifolates, which are among the first antimicrobial agents invented, inhibit cell growth by creating an intracellular state of folate deficiency. Clinical resistance to antifolates has been mainly attributed to mutations that alter structure or expression of enzymes involved in de novo folate synthesis. We identified a Mycobacterium smegmatis mutant, named FUEL (which stands for folate utilization enzyme for leucovorin), that is hypersusceptible to antifolates. Chemical complementation indicated that FUEL is unable to metabolize folinic acid (also known as leucovorin or 5-formyltetrahydrofolate), whose metabolic function remains unknown. Targeted mutagenesis, genetic complementation, and biochemical studies showed that FUEL lacks 5,10-methenyltetrahydrofolate synthase (MTHFS; also called 5-formyltetrahydrofolate cyclo-ligase; EC 6.3.3.2) activity responsible for the only ATP-dependent, irreversible conversion of folinic acid to 5,10-methenyltetrahydrofolate. In trans expression of active MTHFS proteins from bacteria or human restored both antifolate resistance and folinic acid utilization to FUEL. Absence of MTHFS resulted in marked cellular accumulation of polyglutamylated species of folinic acid. Importantly, MTHFS also affected M. smegmatis utilization of monoglutamylated 5-methyltetrahydrofolate exogenously added to the medium. Likewise, Escherichia coli mutants lacking MTHFS became susceptible to antifolates. These results indicate that folinic acid conversion by MTHFS is required for bacterial intrinsic antifolate resistance and folate homeostatic control. This novel mechanism of antimicrobial antifolate resistance might be targeted to sensitize bacterial pathogens to classical antifolates.

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“…Unlike other one-carbon (C 1 ) folates, 5-CHO-THF is not a C 1 donor but a potent inhibitor of many folate-dependent enzymes (Stover and Schirch 1993) and must consequently be removed. 5-FCL is the main enzyme known to do this, and ablating it leads to 5-CHO-THF accumulation (Holmes and Appling 2002; Goyer et al 2005; Jeanguenin et al 2010). 5-FCL is mechanistically a kinase, the initial reaction product being an iminium phosphate intermediate, which then undergoes cyclization and phosphate elimination to give back 5,10-CH=THF (Fig.…”

Section: Introductionmentioning

confidence: 99%

A 5-formyltetrahydrofolate cycloligase paralog from all domains of life: comparative genomic and experimental evidence for a cryptic role in thiamin metabolism

Pribat

Blaby

Lara‐Núñez

et al. 2011

Funct Integr Genomics

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A paralog (here termed COG0212) of the ATP-dependent folate salvage enzyme 5-formyltetrahydrofolate cycloligase (5-FCL) occurs in all domains of life and, although typically annotated as 5-FCL in pro- and eukaryotic genomes, is of unknown function. COG0212 is similar in overall structure to 5-FCL, particularly in the substrate binding region, and has distant similarity to other kinases. The Arabidopsis thaliana COG0212 protein was shown to be targeted to chloroplasts and to be required for embryo viability. Comparative genomic analysis revealed that a high proportion (19%) of archaeal and bacterial COG0212 genes are clustered on the chromosome with various genes implicated in thiamin metabolism or transport but showed no such association between COG0212 and folate metabolism. Consistent with the bioinformatic evidence for a role in thiamin metabolism, ablating COG0212 in the archaeon Haloferax volcanii caused accumulation of thiamin monophosphate. Biochemical and functional complementation tests of several known and hypothetical thiamin-related activities (involving thiamin, its breakdown products, and their phosphates) were, however, negative. Also consistent with the bioinformatic evidence, the COG0212 proteins from A. thaliana and prokaryote sources lacked 5-FCL activity in vitro and did not complement the growth defect or the characteristic 5-formyltetrahydrofolate accumulation of a 5-FCL-deficient (ΔygfA) Escherichia coli strain. We therefore propose (a) that COG0212 has an unrecognized yet sometimes crucial role in thiamin metabolism, most probably in salvage or detoxification, and (b) that is not a 5-FCL and should no longer be so annotated.

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Moonlighting Glutamate Formiminotransferases Can Functionally Replace 5-Formyltetrahydrofolate Cycloligase

Cited by 26 publications

References 59 publications

6S RNA – an old issue became blue-green

6S RNA – an old issue became blue-green

Bacterial Conversion of Folinic Acid Is Required for Antifolate Resistance

A 5-formyltetrahydrofolate cycloligase paralog from all domains of life: comparative genomic and experimental evidence for a cryptic role in thiamin metabolism

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