'LeishMan' topoisomerase I

An ideal chimera for unraveling the role of the small subunit of unusual bi-subunit topoisomerase I from Leishmania donovani

Agneyo Ganguly, Benu Brata Das, Nilkantha Sen, Amit Roy, Somdeb Bose Dasgupta, Hemanta K. Majumder

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The active site tyrosine residue of all monomeric type IB topoisomerases resides in the C-terminal domain of the enzyme. Leishmania donovani, possesses unusual heterodimeric type IB topoisomerase. The small subunit harbors the catalytic tyrosine within the SKXXY motif. To explore the functional relationship between the two subunits, we have replaced the small subunit of L.donovani topoisomerase I with a C-terminal fragment of human topoisomerase I (HTOP14). The purified LdTOP1L (large subunit of L.donovani topoisomerase I) and HTOP14 were able to reconstitute topoisomerase I activity when mixed in vitro. This unusual enzyme, 'LeishMan' topoisomerase I (Leish for Leishmania and Man for human) exhibits less efficiency in DNA binding and strand passage compared with LdTOP1L/S. Fusion of LdTOP1L with HTOP14 yielded a more efficient enzyme with greater affinity for DNA and faster strand passage ability. Both the chimeric enzymes are less sensitive to camptothecin than LdTOP1L/S. Restoration of topoisomerase I activity by LdTOP1L and HTOP14 suggests that the small subunit of L.donovani topoisomerase I is primarily required for supplying the catalytic tyrosine. Moreover, changes in the enzyme properties due to substitution of LdTOP1S with HTOP14 indicate that the small subunit contributes to subunit interaction and catalytic efficiency of the enzyme.

Original languageEnglish (US)
Pages (from-to)6286-6297
Number of pages12
JournalNucleic Acids Research
Volume34
Issue number21
DOIs
StatePublished - Dec 1 2006

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Leishmania donovani
Type I DNA Topoisomerase
Enzymes
Tyrosine
Catalytic Domain
Camptothecin
Leishmania
DNA

ASJC Scopus subject areas

  • Genetics

Cite this

'LeishMan' topoisomerase I : An ideal chimera for unraveling the role of the small subunit of unusual bi-subunit topoisomerase I from Leishmania donovani. / Ganguly, Agneyo; Das, Benu Brata; Sen, Nilkantha; Roy, Amit; Dasgupta, Somdeb Bose; Majumder, Hemanta K.

In: Nucleic Acids Research, Vol. 34, No. 21, 01.12.2006, p. 6286-6297.

Research output: Contribution to journalArticle

Ganguly, Agneyo ; Das, Benu Brata ; Sen, Nilkantha ; Roy, Amit ; Dasgupta, Somdeb Bose ; Majumder, Hemanta K. / 'LeishMan' topoisomerase I : An ideal chimera for unraveling the role of the small subunit of unusual bi-subunit topoisomerase I from Leishmania donovani. In: Nucleic Acids Research. 2006 ; Vol. 34, No. 21. pp. 6286-6297.
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abstract = "The active site tyrosine residue of all monomeric type IB topoisomerases resides in the C-terminal domain of the enzyme. Leishmania donovani, possesses unusual heterodimeric type IB topoisomerase. The small subunit harbors the catalytic tyrosine within the SKXXY motif. To explore the functional relationship between the two subunits, we have replaced the small subunit of L.donovani topoisomerase I with a C-terminal fragment of human topoisomerase I (HTOP14). The purified LdTOP1L (large subunit of L.donovani topoisomerase I) and HTOP14 were able to reconstitute topoisomerase I activity when mixed in vitro. This unusual enzyme, 'LeishMan' topoisomerase I (Leish for Leishmania and Man for human) exhibits less efficiency in DNA binding and strand passage compared with LdTOP1L/S. Fusion of LdTOP1L with HTOP14 yielded a more efficient enzyme with greater affinity for DNA and faster strand passage ability. Both the chimeric enzymes are less sensitive to camptothecin than LdTOP1L/S. Restoration of topoisomerase I activity by LdTOP1L and HTOP14 suggests that the small subunit of L.donovani topoisomerase I is primarily required for supplying the catalytic tyrosine. Moreover, changes in the enzyme properties due to substitution of LdTOP1S with HTOP14 indicate that the small subunit contributes to subunit interaction and catalytic efficiency of the enzyme.",
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