A tyrosyl DNA phosphodiesterase 1 from kinetoplastid parasite Leishmania donovani (LdTdp1) capable of removing topo I-DNA covalent complexes

Bijoylaxmi Banerjee, Amit Roy, Nilkantha Sen, Hemanta K. Majumder

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Tyrosyl DNA phosphodiesterase 1 (Tdp1) is a member of phospholipase D superfamily, which cleaves a broad range of 3′-DNA adducts, the best characterized of which is the phosphodiester bond formed between DNA and topoisomerase IB. This study describes cloning and functional characterization of the enzyme, termed as LdTdp1 in the kinetoplastid parasite Leishmania donovani. Sequence analysis confirmed conservation of the active site motifs typical for all Tdp1 proteins. LdTdp1 activity was detected in the parasite nucleus as well as in the kinetoplast. The enzyme harbours a nuclear localization signal at its C-terminus. Overexpression of the active enzyme protected the parasites against topoisomerase IB inhibitor camptothecin (CPT) and oxidative agent H 2O 2-mediated cytotoxicity and its downregulation rendered the parasites hypersensitive to CPT. Trapping of mutant LdTdp1 on DNA takes place following CPT treatment in L. donovani cells. The expression level and associated activity of LdTdp1 were found to be higher in CPT-resistant L. donovani parasites. Altogether, this is the first report of Tdp1 from the kinetoplastid parasite L. donovani, which actively participates in topoisomerase I-mediated DNA damage repair process and thereby counteracts the cytotoxic effect of topoisomerase I inhibitors.

Original languageEnglish (US)
Pages (from-to)119-137
Number of pages19
JournalMolecular Microbiology
Volume78
Issue number1
DOIs
StatePublished - Oct 1 2010

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Leishmania donovani
Camptothecin
Parasites
DNA
Type I DNA Topoisomerase
Enzymes
Topoisomerase Inhibitors
Topoisomerase I Inhibitors
Nuclear Localization Signals
Phospholipase D
DNA Adducts
DNA Repair
DNA Damage
Sequence Analysis
tyrosyl-DNA phosphodiesterase
Organism Cloning
Catalytic Domain
Down-Regulation
Proteins

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

A tyrosyl DNA phosphodiesterase 1 from kinetoplastid parasite Leishmania donovani (LdTdp1) capable of removing topo I-DNA covalent complexes. / Banerjee, Bijoylaxmi; Roy, Amit; Sen, Nilkantha; Majumder, Hemanta K.

In: Molecular Microbiology, Vol. 78, No. 1, 01.10.2010, p. 119-137.

Research output: Contribution to journalArticle

Banerjee, Bijoylaxmi ; Roy, Amit ; Sen, Nilkantha ; Majumder, Hemanta K. / A tyrosyl DNA phosphodiesterase 1 from kinetoplastid parasite Leishmania donovani (LdTdp1) capable of removing topo I-DNA covalent complexes. In: Molecular Microbiology. 2010 ; Vol. 78, No. 1. pp. 119-137.
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