Conservation of genetic linkage between heat shock protein 100 and glycosylphosphatidylinositol-specific phospholipase C in Trypanosoma brucei and Trypanosoma cruzi

Conservation of genetic linkage between heat shock protein 100 and glycosylphosphatidylinositol-specific phospholipase C in Trypanosoma brucei and Trypanosoma cruzi

Autor Redpath, M. B. Google Scholar
Carnall, N. Google Scholar
Webb, H. Google Scholar
Courel, M. Google Scholar
Amorim, A. Google Scholar
Guther, MLS Google Scholar
Almeida, MLC de Google Scholar
Carrington, M. Google Scholar
Instituição Univ Cambridge
Universidade Federal de São Paulo (UNIFESP)
Univ Dundee
Resumo The experiments described in this paper were designed to try and isolate a recombinant DNA clone encoding a Trypanosoma cruzi homologue of the Trypanosoma br brucei glycosylphosphatidylinositol-specific phospholipase C (GPI-PLC) gene. Despite the ready biochemical detection of phospholipase C activities that hydrolyse GPI-anchors of cell surface proteins in T. cruzi, it did not prove possible to isolate any recombinant DNA clones using the T. brucei gpi-plc gene as a probe. On determining the DNA sequence to the 5' side of the gpi-plc gene it was found to be adjacent to a gene that encodes a 100 kDa heat shock protein (HSP100). To investigate whether this linkage between the hsp100 and gpi-plc genes was conserved in T. cruzi, a probe derived from the T. brucei hsp100 gene was used to isolate T. cruzi genomic clones. These were partially sequenced and shown to contain an hsp100 gene. Restriction enzyme fragments located to the 3' side of the T. cruzi hsp100 gene were then sequenced and found to contain a gene that encodes a polypeptide (TcPLC1) that has 46% amino acid sequence identity with the T. brucei GPI-PLC including most of the key residues involved in inositol binding and the catalytic histidine. A recombinant form of TcPLC1 was produced and shown to possess phospholipase C activity towards a GPI-substrate. Thus, the hsp100 and gpi-plc genes are adjacent in T. brucei and this linkage is conserved in T. cruzi. This observation has been used to facilitate the isolation of a clone encoding a T. cruzi phospholipase C gene. (C) 1998 Published by Elsevier Science B.V. All rights reserved.
Palavra-chave Trypanosoma brucei
Trypanosoma cruzi
glycosylphosphatidylinositol
phospholipase C
heat shock
protein 100
Idioma Inglês
Data de publicação 1998-07-01
Publicado em Molecular and Biochemical Parasitology. Amsterdam: Elsevier B.V., v. 94, n. 1, p. 113-121, 1998.
ISSN 0166-6851 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 113-121
Fonte http://dx.doi.org/10.1016/S0166-6851(98)00056-5
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000075308600009
Endereço permanente http://repositorio.unifesp.br/handle/11600/25926

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