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Analysis of NLS and rRNA binding motifs in the L25 ribosomal protein from Leishmania (Viannia) braziliensis: investigation of its diagnostic capabilities

Published online by Cambridge University Press:  16 January 2003

A. C. GONZÁLEZ ,
E. MARTÍNEZ ,
E. CARMELO ,
J. E. PIÑERO ,
V. ALONSO ,
A. DEL CASTILLO  and
B. VALLADARES
A. C. GONZÁLEZ
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
E. MARTÍNEZ
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
E. CARMELO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
J. E. PIÑERO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
V. ALONSO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
A. DEL CASTILLO
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
B. VALLADARES
Affiliation:
Department of Parasitology, Faculty of Pharmacy, University of La Laguna, Avda. Francisco Sánchez s/n, CP38271 La Laguna, Tenerife, Spain
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Abstract

A cDNA clone codifying ribosomal protein L25 was isolated from a Leishmania braziliensis cDNA gene library. The alignment of the amino acid sequence deduced from this gene with other proteins revealed that this protein is related to the L23/25 ribosomal protein family. This is so because this protein shows, in its C-terminal end, the rRNA binding domains characteristic of these proteins and at the N-terminal end the NLS sequence necessary for its entry into the nucleus. Southern blot analysis showed 2 copies of gene L25 per genome arranged in tandem position and pointing in the same direction. Northern blot analysis showed that this gene is transcribed in 2 mRNAs when parasite promastigotes are in the logarithmic phase. In order to analyse the antigenic properties of L. braziliensis RPL25, it was purified as a recombinant protein and ELISA-tested against cutaneous, mucocutaneous and Chagasic sera. The results indicate that the recombinant RPL25 from L. braziliensis presents a non-specific reaction that disqualifies it for the diagnosis of cutaneous leishmaniasis. In contrast, some of the synthetic peptides derived from its sequence may serve as promising tools for the diagnosis of this disease.

Keywords

ribosomal protein L25Leishmania braziliensisnuclear localisation sequence (NLS)rRNA binding motifs
Type
Research Article
Information
Parasitology , Volume 125 , Issue 1 , July 2002 , pp. 51 - 57
Copyright
2002 Cambridge University Press

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