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The study of genomic variability within various pathogenic and non-pathogenic strains of mycobacteria provides insight into their evolution and pathogenesis. The mycobacterial genome encodes seven cutinase-like proteins and each one of these exhibit distinct characteristics. We describe the presence of Cut5, a member of the cutinase family, in mycobacteria and the existence of a unique genomic arrangement in the  cut5  gene of  M .  tuberculosis  ( Mtb ) strains. A single nucleotide (T) insertion is observed in the  cut5  gene, which is specific for  Mtb  strains. Using  in silico  analysis and RT-PCR, we demonstrate the transcription of  Rv3724/cut5  as  Rv3724a/cut5a  and  Rv3724b/cut5b  in  Mtb  H37Rv and as full length  cut5  in  M .  bovis . Cut5b protein of  Mtb  H37Rv ( Mtb Cut5b) was found to be antigenically similar to its homologs in  M .  bovis  and  M .  smegmatis , without any observed cross-reactivity with other  Mtb  cutinases. Also, the presence of Cut5b in  Mtb  and its homologs in  M .  bovis  and  M .  smegmatis  were confirmed by western blotting using antibodies raised against recombinant Cut5b. In  Mtb  H37Rv, Cut5b was found to be localized in the cell wall, cytosol and membrane fractions. We also report the vast prevalence of Cut5 homologs in pathogenic and non pathogenic species of mycobacteria.  In silico  analysis revealed that this protein has three possible organizations in mycobacteria. Also, a single nucleotide (T) insertion in  Mtb  strains and varied genomic arrangements within mycobacterial species make Rv3724/Cut5 a potential candidate that can be exploited as a biomarker in  Mtb  infection.

Heterogeneity among Homologs of Cutinase-Like Protein Cut5 in Mycobacteria