Insights from the Structure of Mycobacterium tuberculosis Topoisomerase i with a Novel Protein Fold

Kemin Tan, Nan Cao, Bokun Cheng, Andrzej Joachimiak, Yuk Ching Tse-Dinh

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The DNA topoisomerase I enzyme of Mycobacterium tuberculosis (MtTOP1) is essential for the viability of the organism and survival in a murine model. This topoisomerase is being pursued as a novel target for the discovery of new therapeutic agents for the treatment of drug-resistant tuberculosis. In this study, we succeeded in obtaining a structure of MtTOP1 by first predicting that the C-terminal region of MtTOP1 contains four repeated domains that do not involve the Zn-binding tetracysteine motifs seen in the C-terminal domains of Escherichia coli topoisomerase I. A construct (amino acids A2-T704), MtTOP1-704t, that includes the N-terminal domains (D1-D4) and the first predicted C-terminal domain (D5) of MtTOP1 was expressed and found to retain DNA cleavage-religation activity and catalyze single-stranded DNA catenation. MtTOP1-704t was crystallized, and a structure of 2.52 Å resolution limit was obtained. The structure of the MtTOP1 N-terminal domains has features that have not been observed in other previously available bacterial topoisomerase I crystal structures. The first C-terminal domain D5 forms a novel protein fold of a four-stranded antiparallel β-sheet stabilized by a crossing-over α-helix. Since there is only one type IA topoisomerase present in Mycobacteriaceae and related Actinobacteria, this subfamily of type IA topoisomerase may be required for multiple functions in DNA replication, transcription, recombination, and repair. The unique structural features observed for MtTOP1 may allow these topoisomerase I enzymes to carry out physiological functions associated with topoisomerase III enzyme in other bacteria.

Original languageEnglish (US)
Pages (from-to)182-193
Number of pages12
JournalJournal of Molecular Biology
Volume428
Issue number1
DOIs
StatePublished - Jan 16 2016
Externally publishedYes

Fingerprint

Type I DNA Topoisomerase
Mycobacterium tuberculosis
Mycobacteriaceae
Proteins
Enzymes
Recombinational DNA Repair
Multidrug-Resistant Tuberculosis
DNA Cleavage
Actinobacteria
Single-Stranded DNA
DNA Replication
varespladib methyl
Escherichia coli
Bacteria
Amino Acids
Therapeutics

Keywords

  • crystallization
  • protein domains
  • protein fold
  • TB
  • TopA

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

Cite this

Insights from the Structure of Mycobacterium tuberculosis Topoisomerase i with a Novel Protein Fold. / Tan, Kemin; Cao, Nan; Cheng, Bokun; Joachimiak, Andrzej; Tse-Dinh, Yuk Ching.

In: Journal of Molecular Biology, Vol. 428, No. 1, 16.01.2016, p. 182-193.

Research output: Contribution to journalArticle

Tan, Kemin ; Cao, Nan ; Cheng, Bokun ; Joachimiak, Andrzej ; Tse-Dinh, Yuk Ching. / Insights from the Structure of Mycobacterium tuberculosis Topoisomerase i with a Novel Protein Fold. In: Journal of Molecular Biology. 2016 ; Vol. 428, No. 1. pp. 182-193.
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