Plasmodium serine hydroxymethyltransferase: indispensability and display of distinct localization

Malaria Journal

Research

Wichai Pornthanakasem¹  Chairat Uthaipibull¹  Ubolsree Leartsakulpanich¹  Darin Kongkasuriyachai¹  Yongyuth Yuthavong¹ 

[1] National Center for Genetic Engineering and Biotechnology, 113 Phahonyothin Road, Khlong Nueng, 12120, Pathum Thani, Khlong Luang, Thailand;

关键词: Plasmodium;    Serine hydroxymethyltransferase;    Localization;   

DOI  :  10.1186/1475-2875-11-387

 received in 2012-08-28, accepted in 2012-11-20,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundSerine hydroxymethyltransferase (SHMT), a pyridoxal phosphate-dependent enzyme, plays a vital role in the de novo pyrimidine biosynthesis pathway in malaria parasites. Two genes have been identified in Plasmodium spp. encoding a cytosolic SHMT (cSHMT) and putative mitochondria SHMT (mSHMT), but their roles have not been fully investigated.MethodsThe presence of Plasmodium SHMT isoforms in the intra-erythrocytic stage was assessed based on their gene expression using reverse transcription PCR (RT-PCR). Localization studies of Plasmodium SHMT isoforms were performed by transfection of fluorescent-tagged gene constructs into P. falciparum and expressions of fluorescent fusion proteins in parasites were observed using a laser scanning confocal microscope. Genetic targeting through homologous recombination was used to study the essentiality of SHMT in Plasmodium spp.ResultsSemi-quantitative RT-PCR revealed the expression of these two genes throughout intra-erythrocytic development. Localization studies using P. falciparum expressing fluorescent-tagged SHMT showed that Pf cSHMT-red fluorescent fusion protein (Pf cSHMT-DsRed) is localized in the cytoplasm, while Pf mSHMT-green fluorescent fusion protein (Pf mSHMT-GFP) co-localized with Mitotracker™-labelled mitochondria as predicted. The essentiality of plasmodial cSHMT was inferred from transfection experiments where recovery of viable knock-out parasites was not achieved, unless complemented with a functional equivalent copy of shmt.ConclusionsDistinct compartment localizations of Pf SHMT were observed between cytoplasmic and mitochondrial isoforms, and evidence was provided for the indispensable role of plasmodial cSHMT indicating it as a valid target for development of novel anti-malarials.

【 授权许可】

Unknown   
© Pornthanakasem et al.; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.