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Characterisation of the Gene Encoding type II DNA Topoisomerase from Leishmania Donovani: a Key Molecular Target in Antileishmanial Therapy

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Title Characterisation of the Gene Encoding type II DNA Topoisomerase from Leishmania Donovani: a Key Molecular Target in Antileishmanial Therapy
 
Creator Das, Aditi
Dasgupta, Arindam
Sharma, Shalini
Ghosh, Monidipa
Sengupta, Tanusree
Bandyopadhyay, Santu
Majumder, Hemanta K
 
Subject Infectious Diseases and Immunology
 
Description The gene encoding type II DNA topoisomerase from the kinetoplastid hemoflagellated protozoan parasite Leishmania donovani (LdTOP2) was isolated from a genomic DNA library of this parasite. DNA sequence analysis revealed an ORF of 3711 bp encoding a putative protein of 1236 amino acids with no introns. The deduced amino acid sequence of LdTOP2 showed strong homologies to TOP2 sequences from other kinetoplastids, namely Crithidia and Trypanosoma spp. with estimated identities of 86 and 68%, respectively. LdTOP2 shares a much lower identity of 32% with its human homologue. LdTOP2 is located as a single copy on a chromosome in the 0.7 Mb region in the L.donovani genome and is expressed as a 5 kb transcript. 5′-Mapping studies indicate that the LdTOP2 gene transcript is matured post-transcriptionally with the trans-splicing of the mini-exon occurring at –639 from the predicted initiation site. Antiserum raised in rabbit against glutathione S-transferase fusion protein containing the major catalytic portion of the recombinant L.donovani topoisomerase II protein could detect a band on western blots at ∼132 kDa, the expected size of the entire protein. Use of the same antiserum for immunolocalisation analysis led to the identification of nuclear, as well as kinetoplast, antigens for L.donovani topoisomerase II. The in vitro biochemical properties of the full-length recombinant LdTOP2 when overexpressed in E.coli were similar to the Mg(II) and ATP-dependent activity found in cell extracts of L.donovani.
 
Publisher Oxford University Press
 
Date 2001
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://www.eprints.iicb.res.in/1642/1/Nucleic_Acids_Research%2C_2001.pdf
Das, Aditi and Dasgupta, Arindam and Sharma, Shalini and Ghosh, Monidipa and Sengupta, Tanusree and Bandyopadhyay, Santu and Majumder, Hemanta K (2001) Characterisation of the Gene Encoding type II DNA Topoisomerase from Leishmania Donovani: a Key Molecular Target in Antileishmanial Therapy. Nucleic Acids Research, 29 (9). pp. 1844-1851.
 
Relation http://dx.doi.org/10.1093/nar/29.9.1844
http://www.eprints.iicb.res.in/1642/