Entamoeba histolytica: differential gene expression during programmed cell death and identification of early pro- and anti-apoptotic signals.
We have demonstrated that programmed cell death (PCD) in Entamoeba histolytica is induced in vitro by G418 aminoglycoside antibiotic. To ascertain if biochemical and morphological changes previously observed are paired to molecular changes that reflect a genetic program, we looked here for early differential gene expression during the induction of PCD. Using cDNA-amplified fragment length polymorphisms (AFLPs) and in silico derived analysis we showed in E. histolytica a differential gene expression during PCD induced by G418. The genes identified encoded for proteins homologous to Glutaminyl-tRNA synthase, Ribosomal Subunit Proteins 40S and 18S, Saposin-like, Silent Information Regulator-2 (Sir-2), and Grainins 1 and 2. Using real-time quantitative PCR (RT Q-PCR), we found that glutaminyl-tRNA synthetase, sir-2, grainins and saposin-like genes were strongly overexpressed after 30min of PCD induction, while its expression dramatically decreased up to 60min. On the other hand, overexpression of ribosomal genes increased only 7-fold of basal expression, showing a progressive down-regulation up to 90min. glutaminyl-tRNA synthetase, sir-2 and grainins could act as negative regulators of PCD, trying to control the biochemical changes related to PCD activation. Overexpression of saposin-like gene could act as up-regulator of some cell death pathways. Our results give evidence of the first genes identified during the early stage of PCD in E. histolytica that could be implicated in regulation of apoptotic pathways.
Laboratorio de Biomedicina Molecular I, Programa Institucional de Biomedicina Molecular, Escuela Nacional de Medicina y Homeopatía, IPN, Guillermo Massieu Helguera No. 239, Fracc. La Escalera, Col. Ticomán, México, DF, México.
Virginia Sánchez
VS
Monroy
Ma Olivia Medel
MO
Flores
José D'Artagnan
JD
Villalba-Magdaleno
Consuelo Gómez
CG
Garcia
David Guillermo Pérez
DG
Ishiwara
D000563:Amebicides / Q000494:pharmacology; D000595:Amino Acid Sequence; D000604:Amino Acyl-tRNA Synthetases / Q000096:biosynthesis / Q000235:genetics; D054458:Amplified Fragment Length Polymorphism Analysis; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D002135:Calcium-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry; D004748:Entamoeba histolytica / Q000166:cytology / Q000187:drug effects / Q000235:genetics*; D015870:Gene Expression; D005839:Gentamicins / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000379:methods; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics; D049231:Saposins / Q000096:biosynthesis / Q000235:genetics; D016415:Sequence Alignment; D037761:Sirtuins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
D000563:Amebicides; D002135:Calcium-Binding Proteins; D018076:DNA, Complementary; D005839:Gentamicins; D015800:Protozoan Proteins; D049231:Saposins; C418423:grainin 1 protein, Entamoeba histolytica; C418424:grainin 2 protein, Entamoeba histolytica; C010680:antibiotic G 418; D037761:Sirtuins; D000604:Amino Acyl-tRNA Synthetases; C020659:glutaminyl-tRNA synthetase