TY - JOUR
T1 - Entamoeba histolytica
T2 - Differential gene expression during programmed cell death and identification of early pro- and anti-apoptotic signals
AU - Monroy, Virginia Sánchez
AU - Flores, Ma Olivia Medel
AU - Villalba-Magdaleno, José D.Artagnan
AU - Garcia, Consuelo Gómez
AU - Ishiwara, David Guillermo Pérez
N1 - Funding Information:
This work was supported by CONACYT and IPN Grants given to D.G.P.I. The authors express our gratitude to Dr. Kevin G. McGuigan, RCSI Irish for critical reading.
PY - 2010/12
Y1 - 2010/12
N2 - 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 30. min of PCD induction, while its expression dramatically decreased up to 60. min. On the other hand, overexpression of ribosomal genes increased only 7-fold of basal expression, showing a progressive down-regulation up to 90. min. g. lutaminyl-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.
AB - 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 30. min of PCD induction, while its expression dramatically decreased up to 60. min. On the other hand, overexpression of ribosomal genes increased only 7-fold of basal expression, showing a progressive down-regulation up to 90. min. g. lutaminyl-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.
KW - Apoptosis
KW - CDNA-AFLPs
KW - E. histolytica
KW - Gene expression
KW - PCD
UR - http://www.scopus.com/inward/record.url?scp=79955764641&partnerID=8YFLogxK
U2 - 10.1016/j.exppara.2010.05.027
DO - 10.1016/j.exppara.2010.05.027
M3 - Artículo
SN - 0014-4894
VL - 126
SP - 497
EP - 505
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 4
ER -