Expression and cellular localization of Trypanosoma cruzi type II DNA topoisomerase.

Abstract:

:Topoisomerases are enzymes that participate in many cellular functions involving topological manipulation of DNA strands. There are two types of topoisomerases in the cell: (a) type I topoisomerases; and (b) type II topoisomerases (topo II). Previously we have cloned and sequenced the gene encoding Trypanosoma cruzi topo II (TcTOP2). This study group has raised an antiserum against recombinant type II DNA topoisomerase (TctopoII) to study the expression of this gene during T. cruzi differentiation and to determine the cellular location of the enzyme. Western blot analysis showed that T. cruzi TctopoII is expressed in the replicative epimastigotes but not in the infective and non-replicative trypomastigotes. However, slot blot analysis of RNAs extracted from epimastigotes and metacyclic trypomastigotes showed that the mRNA encoding the enzyme is present in both developmental stages of the parasite. Confocal laser microscopy using the antiserum raised against recombinant TctopoII showed that the enzyme is located exclusively in the nucleus of the parasite. Similar results were obtained by immunofluorescence analysis of Crithidia fasciculata. However, monoclonal antisera against the corresponding enzyme extracted from C. fasciculata recognizes a kinetoplast protein in both T. cruzi and Crithidia.

journal_name

Mol Biochem Parasitol

authors

Fragoso SP,Mattei D,Hines JC,Ray D,Goldenberg S

doi

10.1016/s0166-6851(98)00066-8

subject

Has Abstract

pub_date

1998-08-01 00:00:00

pages

197-204

issue

2

eissn

0166-6851

issn

1872-9428

pii

S0166-6851(98)00066-8

journal_volume

94

pub_type

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