Engineering a self-sufficient Mycobacterium tuberculosis CYP130 by gene fusion with the reductase-domain of CYP102A1 from Bacillus megaterium

Abstract Mycobacterium tuberculosis is a facultative intracellular pathogen responsible for causing tuberculosis. The harsh environment in which M. tuberculosis survives requires this pathogen to continuously adapt in order to maintain an evolutionary advantage. However, the apparent absence of horizontal gene transfer in M. tuberculosis imposes restrictions in the ways by which evolution can occur. Large-scale changes in the genome can be introduced through genome reduction, recombination events and structural variation. Here, we identify a functional chimeric protein in the ppe38–71 locus, the absence of which is known to have an impact on protein secretion and virulence. To examine whether this approach was used more often by this pathogen, we further develop software that detects potential gene fusion events from multigene deletions using whole genome sequencing data. With this software we could identify a number of other putative gene fusion events within the genomes of M. tuberculosis isolates. We were able to demonstrate the expression of one of these gene fusions at the protein level using mass spectrometry. Therefore, gene fusions may provide an additional means of evolution for M. tuberculosis in its natural environment whereby novel chimeric proteins and functions can arise.

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Electron Microscopic Observations on the Sites of Activities of Succinic Dehydrogenase and Cytochrome Oxidase in Mycobacterium tuberculosis and Bacillus megaterium

Journal of Electron Microscopy ◽

10.1093/oxfordjournals.jmicro.a049533 ◽

1966 ◽

Keyword(s):

Mycobacterium Tuberculosis ◽

Cytochrome Oxidase ◽

Bacillus Megaterium ◽

Succinic Dehydrogenase ◽

Electron Microscopic

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Bacillus halodurans Strain C125 Encodes and Synthesizes Enzymes from Both Known Pathways To Form dUMP Directly from Cytosine Deoxyribonucleotides

Applied and Environmental Microbiology ◽

10.1128/aem.00268-15 ◽

2015 ◽

Vol 81 (10) ◽

pp. 3395-3404 ◽

Cited By ~ 1

Author(s):

Christian Berg Oehlenschlæger ◽

Monika Nøhr Løvgreen ◽

Eva Reinauer ◽

Emilia Lehtinen ◽

Marie-Louise Lindberg Pind ◽

...

Keyword(s):

Escherichia Coli ◽

Mycobacterium Tuberculosis ◽

Bacillus Thuringiensis ◽

Bacillus Megaterium ◽

Cytidine Deaminase ◽

Bacillus Halodurans ◽

Whole Genome ◽

Gene Encoding ◽

Content Type ◽

Regulatory Properties

ABSTRACTAnalysis of the genome ofBacillus haloduransstrain C125 indicated that two pathways leading from a cytosine deoxyribonucleotide to dUMP, used for dTMP synthesis, were encoded by the genome of the bacterium. The genes that were responsible, thecomEBgene and thedcdBgene, encoding dCMP deaminase and the bifunctional dCTP deaminase:dUTPase (DCD:DUT), respectively, were both shown to be expressed inB. halodurans, and both genes were subject to repression by the nucleosides thymidine and deoxycytidine. The latter nucleoside presumably exerts its repression after deamination by cytidine deaminase. BothcomEBanddcdBwere cloned, overexpressed inEscherichia coli, and purified to homogeneity. Both enzymes were active and displayed the expected regulatory properties: activation by dCTP for dCMP deaminase and dTTP inhibition for both enzymes. Structurally, theB. haloduransenzyme resembled theMycobacterium tuberculosisenzyme the most. An investigation of sequenced genomes from other species of the genusBacillusrevealed that not only the genome ofB. haloduransbut also the genomes ofBacillus pseudofirmus,Bacillus thuringiensis,Bacillus hemicellulosilyticus,Bacillus marmarensis,Bacillus cereus, andBacillus megateriumencode both the dCMP deaminase and the DCD:DUT enzymes. In addition, eightdcdBhomologs fromBacillusspecies within the genus for which the whole genome has not yet been sequenced were registered in the NCBI Entrez database.

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