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Literature summary for 5.6.2.1 extracted from

  • Narula, G.; Becker, J.; Cheng, B.; Dani, N.; Abrenica, M.V.; Tse-Dinh, Y.C.
    The DNA relaxation activity and covalent complex accumulation of Mycobacterium tuberculosis topoisomerase I can be assayed in Escherichia coli: application for identification of potential FRET-dye labeling sites (2010), BMC Biochem., 11, 41.
    View publication on PubMedView publication on EuropePMC

Application

Application Comment Organism
analysis the absence of cysteine residues in MtTOP1 makes it an attractive system for introduction of potentially informative chemical or spectroscopic probes at specific positions via cysteine mutagenesis. Such probes could be useful for development of high throughput screening assays and can be utilized for fluorescence probe incorporation and fluorescence resonance energy transfer measurement with fluorophore-labeled oligonucleotide substrate Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment) Organism
plasmid-encoded recombinant MtTOP1 can complement the temperature sensitive topA function of Escherichia coli strain AS17 for growth at 42°C. The relaxation activity of recombinant wild-type MtTOP1 is functional in Escherichia coli. No complementation from the Y342A or G116S MtTOP1 mutants, but by mutant H139C, L170C, Y174C, T142C, and K524C. Overexpression of MtTOP1 mutant G116S leads to accumulation of cleavage complex formed by MtTOP1 in Escherichia coli Mycobacterium tuberculosis

Protein Variants

Protein Variants Comment Organism
D111N site-directed mutagenesis, the mutation is lethal for the host Mycobacterium tuberculosis
G116S site-directed mutagenesis, MtTOP1 TOPRIM motif mutation, the mutation inhibits DNA religation. The DNA cleavage activity of MtTOP1-G116 S is Mg2+-dependent Mycobacterium tuberculosis
H139C site-directed mutagenesis, the mutant can be labeled with fluorophores with no significant loss of relaxation activity, the mutant complements the temperature sensitive topA function of Escherichia coli strain AS17 Mycobacterium tuberculosis
K524C site-directed mutagenesis, the mutant can be labeled with fluorophores with no significant loss of relaxation activity, the mutant complements the temperature sensitive topA function of Escherichia coli strain AS17 Mycobacterium tuberculosis
L170C site-directed mutagenesis, the mutant can be labeled with fluorophores with no significant loss of relaxation activity, the mutant complements the temperature sensitive topA function of Escherichia coli strain AS17 Mycobacterium tuberculosis
T142C site-directed mutagenesis, the mutant can be labeled with fluorophores with no significant loss of relaxation activity, the mutant complements the temperature sensitive topA function of Escherichia coli strain AS17 Mycobacterium tuberculosis
Y174C site-directed mutagenesis, the mutant can be labeled with fluorophores with no significant loss of relaxation activity, the mutant complements the temperature sensitive topA function of Escherichia coli strain AS17 Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ the DNA cleavage activity of MtTOP1-G116 S is Mg2+-dependent Mycobacterium tuberculosis

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
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-
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information structure-function analysis of MtTOP1, overview Mycobacterium tuberculosis ?
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?

Subunits

Subunits Comment Organism
More structure-function analysis of MtTOP1, overview Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
MtTOP1
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Mycobacterium tuberculosis
Topoisomerase I
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Mycobacterium tuberculosis

General Information

General Information Comment Organism
malfunction accumulation of cleavage complex formed by inactivated MtTOP1 is lethal for the cell Mycobacterium tuberculosis
additional information the TOPRIM motif DxDxxG is strictly conserved in type IA topoisomerase sequences and plays an essential role for divalent ion coordination and cleavage-religation of DNA during catalysis Mycobacterium tuberculosis