TY - JOUR
T1 - Multinucleation and Polykaryon Formation is Promoted by the EhPC4 Transcription Factor in Entamoeba histolytica
AU - Cruz, Olga Hernández De La
AU - Marchat, Laurence A.
AU - Guillén, Nancy
AU - Weber, Christian
AU - Rosas, Itzel López
AU - DÍaz-Chávez, José
AU - Herrera, Luis
AU - Rojo-Domínguez, Arturo
AU - Orozco, Esther
AU - López-Camarillo, César
N1 - Funding Information:
We thank Universidad Autonoma de la Ciudad de Mexico (UACM) for support. This work was also supported by CONACYT (59481 grant) and Mexico-France program (SEP-CONACYT-ANUIES; ECOS NORD grant M08-S02). Olga Hernández de la Cruz was a scholarship recipient from ECOS-NORD Mexico-France grant M08-S02 (2008–2012). We thank Tomoyoshi Nozaki for provide us the pKT-3M plasmid, to the Genopole platform from Pasteur Institute Paris for DNA microarray analysis, and to Dr. Rosaura Hernández for provide us the anti-Lamin B1 antibodies.
PY - 2016/1/21
Y1 - 2016/1/21
N2 - Entamoeba histolytica is the intestinal parasite responsible for human amoebiasis that is a leading cause of death in developing countries. In this protozoan, heterogeneity in DNA content, polyploidy and genome plasticity have been associated to alterations in mechanisms controlling DNA replication and cell division. Studying the function of the transcription factor EhPC4, we unexpectedly found that it is functionally related to DNA replication, and multinucleation. Site-directed mutagenesis on the FRFPKG motif revealed that the K 127 residue is required for efficient EhPC4 DNA-binding activity. Remarkably, overexpression of EhPC4 significantly increased cell proliferation, DNA replication and DNA content of trophozoites. A dramatically increase in cell size resulting in the formation of giant multinucleated trophozoites (polykaryon) was also found. Multinucleation event was associated to cytokinesis failure leading to abortion of ongoing cell division. Consistently, genome-wide profiling of EhPC4 overexpressing trophozoites revealed the up-regulation of genes involved in carbohydrates and nucleic acids metabolism, chromosome segregation and cytokinesis. Forced overexpression of one of these genes, EhNUDC (nuclear movement protein), led to alterations in cytokinesis and partially recapitulated the multinucleation phenotype. These data indicate for the first time that EhPC4 is associated with events related to polyploidy and genome stability in E. histolytica.
AB - Entamoeba histolytica is the intestinal parasite responsible for human amoebiasis that is a leading cause of death in developing countries. In this protozoan, heterogeneity in DNA content, polyploidy and genome plasticity have been associated to alterations in mechanisms controlling DNA replication and cell division. Studying the function of the transcription factor EhPC4, we unexpectedly found that it is functionally related to DNA replication, and multinucleation. Site-directed mutagenesis on the FRFPKG motif revealed that the K 127 residue is required for efficient EhPC4 DNA-binding activity. Remarkably, overexpression of EhPC4 significantly increased cell proliferation, DNA replication and DNA content of trophozoites. A dramatically increase in cell size resulting in the formation of giant multinucleated trophozoites (polykaryon) was also found. Multinucleation event was associated to cytokinesis failure leading to abortion of ongoing cell division. Consistently, genome-wide profiling of EhPC4 overexpressing trophozoites revealed the up-regulation of genes involved in carbohydrates and nucleic acids metabolism, chromosome segregation and cytokinesis. Forced overexpression of one of these genes, EhNUDC (nuclear movement protein), led to alterations in cytokinesis and partially recapitulated the multinucleation phenotype. These data indicate for the first time that EhPC4 is associated with events related to polyploidy and genome stability in E. histolytica.
UR - http://www.scopus.com/inward/record.url?scp=84955606918&partnerID=8YFLogxK
U2 - 10.1038/srep19611
DO - 10.1038/srep19611
M3 - Artículo
C2 - 26792358
SN - 2045-2322
VL - 6
JO - Scientific Reports
JF - Scientific Reports
M1 - 19611
ER -