TY - JOUR
T1 - Entamoeba histolytica
T2 - Cloning and expression of the poly(A) polymerase EhPAP
AU - García-Vivas, Jessica
AU - López-Camarillo, César
AU - Azuara-Liceaga, Elisa
AU - Orozco, Esther
AU - Marchat, Laurence A.
N1 - Funding Information:
Preliminary EhPap gene sequence data were obtained from TIGR and Sanger E. histolytica genome sequence projects databases. The E. histolytica sequencing effort is part of the Genome Sequencing Project and is supported by awards from The National Institute of Allergy and Infectious Diseases, National Institute of Health, and from The Wellcome Trust. This work was supported by CGPI-IPN (20040056) and CONACyT-México (39888-M). The authors thank Mr. Alfredo Padilla-Barberi for artwork.
PY - 2005/7
Y1 - 2005/7
N2 - In eukaryotes, polyadenylation of pre-mRNA 3′ end is essential for mRNA export, stability, and translation. Here we identified and cloned a gene codifying for a putative nuclear poly(A) polymerase (EhPAP) in Entamoeba histolytica. Protein sequence alignments with eukaryotic PAPs showed that EhPAP has the RNA-binding region and the PAP central domain with the catalytic nucleotidyl transferase domain described for other nuclear PAPs. Recombinant EhPAP expressed in bacteria was used to generate specific antibodies, which recognized two EhPAP isoforms of 60 and 63 kDa in nuclear and cytoplasmic extracts by Western blot assays. RT-PCR assays showed that EhPap mRNA expression varies in multidrug-resistant trophozoites growing in different emetine concentrations. Moreover, EhPap mRNA expression is about 10- and 7-fold increased in G1 and S phase, respectively, through cell cycle progression. These results suggest the existence of a link between EhPAP expression and MDR and cell cycle regulation, respectively.
AB - In eukaryotes, polyadenylation of pre-mRNA 3′ end is essential for mRNA export, stability, and translation. Here we identified and cloned a gene codifying for a putative nuclear poly(A) polymerase (EhPAP) in Entamoeba histolytica. Protein sequence alignments with eukaryotic PAPs showed that EhPAP has the RNA-binding region and the PAP central domain with the catalytic nucleotidyl transferase domain described for other nuclear PAPs. Recombinant EhPAP expressed in bacteria was used to generate specific antibodies, which recognized two EhPAP isoforms of 60 and 63 kDa in nuclear and cytoplasmic extracts by Western blot assays. RT-PCR assays showed that EhPap mRNA expression varies in multidrug-resistant trophozoites growing in different emetine concentrations. Moreover, EhPap mRNA expression is about 10- and 7-fold increased in G1 and S phase, respectively, through cell cycle progression. These results suggest the existence of a link between EhPAP expression and MDR and cell cycle regulation, respectively.
KW - Entamoeba histolytica
KW - Poly(A) polymerase
KW - Polyadenylation
KW - Pre-mRNA 3′ end processing
UR - http://www.scopus.com/inward/record.url?scp=20444504679&partnerID=8YFLogxK
U2 - 10.1016/j.exppara.2005.02.017
DO - 10.1016/j.exppara.2005.02.017
M3 - Artículo
C2 - 15955317
AN - SCOPUS:20444504679
SN - 0014-4894
VL - 110
SP - 226
EP - 232
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 3 SPEC. ISS.
ER -
