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Related Experiment Videos

Studies on transfer RNA from mycobacteria

D N Deobagkar, K P Gopinathan

    Canadian Journal of Microbiology
    |June 1, 1978
    PubMed
    Summary

    Mycobacterium tRNA and aminoacyl-tRNA synthetases are active in younger cells but less so in older cells. These enzymes efficiently aminoacylate bacterial tRNA but not eukaryotic tRNA.

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    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Microbiology

    Background:

    • Transfer RNA (tRNA) and aminoacyl-tRNA synthetases are crucial for protein synthesis.
    • Investigating these components in mycobacteria is important for understanding their unique biology.

    Purpose of the Study:

    • To isolate and characterize active tRNA and aminoacyl-tRNA synthetases from Mycobacterium smegmatis and Mycobacterium tuberculosis.
    • To assess the substrate specificity and activity of these enzymes.

    Main Methods:

    • Isolation of tRNA and aminoacyl-tRNA synthetases from exponentially growing and older mycobacterial cells.
    • Enzyme assays for aminoacylation using homologous and heterologous tRNAs.
    • Column chromatography for tRNA purification.

    Main Results:

    • Active tRNA and aminoacyl-tRNA synthetases were isolated from M. smegmatis and M. tuberculosis.
    • Enzyme activity was retained in older cells, but tRNA showed reduced activity.
    • Mycobacterial synthetases efficiently aminoacylated bacterial tRNA (M. smegmatis, M. tuberculosis, E. coli) but not eukaryotic tRNA (yeast, rat liver).
    • Specific tRNAs (valine, methionine) were purified, and one tRNAMet species was formylated.

    Conclusions:

    • Mycobacterial aminoacyl-tRNA synthetases exhibit broad specificity for bacterial tRNAs.
    • Age-related modifications affect tRNA activity in mycobacteria.
    • These findings provide insights into mycobacterial translation machinery.

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