title: Multinucleation and Polykaryon Formation is Promoted by the EhPC4 Transcription Factor in Entamoeba histolytica.
abstract: 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 K127 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.
authors.affiliation: Universidad Autonoma de la Ciudad de Mexico, Genomics Sciences Program, Mexico City, Mexico.
authors.forename: Olga
authors.initials: O
authors.lastname: Hernández de la Cruz
authors.affiliation: National Polytechnic Institute, National School of Medicine and Homeopathy, Institutional Program of Molecular Biomedicine, Biotechnology Program, Mexico City, Mexico.
authors.forename: Laurence A
authors.initials: LA
authors.lastname: Marchat
authors.affiliation: Institut Pasteur, Cellular Biology of Parasitism Unit, Paris, France.|INSERM U786, Paris, France.
authors.forename: Nancy
authors.initials: N
authors.lastname: Guillén
authors.affiliation: Institut Pasteur, Cellular Biology of Parasitism Unit, Paris, France.|INSERM U786, Paris, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Weber
authors.affiliation: Universidad Autonoma de la Ciudad de Mexico, Genomics Sciences Program, Mexico City, Mexico.
authors.forename: Itzel
authors.initials: I
authors.lastname: López Rosas
authors.affiliation: National Institute of Cancerology, Carcinogenesis Laboratory, Mexico City, Mexico.
authors.forename: José
authors.initials: J
authors.lastname: Díaz-Chávez
authors.affiliation: National Institute of Cancerology, Carcinogenesis Laboratory, Mexico City, Mexico.
authors.forename: Luis
authors.initials: L
authors.lastname: Herrera
authors.affiliation: Metropolitan Autonomous University, Natural Sciences Department, Mexico City, Mexico.
authors.forename: Arturo
authors.initials: A
authors.lastname: Rojo-Domínguez
authors.affiliation: Center for Research and Advanced Studies of the National Polytechnic Institute, Department of Infectomics and Molecular Pathogenesis, Mexico City, Mexico.
authors.forename: Esther
authors.initials: E
authors.lastname: Orozco
authors.affiliation: Universidad Autonoma de la Ciudad de Mexico, Genomics Sciences Program, Mexico City, Mexico.
authors.forename: César
authors.initials: C
authors.lastname: López-Camarillo
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001665:Binding Sites; D049109:Cell Proliferation; D020090:Chromosome Segregation; D017124:Conserved Sequence; D048749:Cytokinesis; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication; D004748:Entamoeba histolytica / Q000235:genetics* / Q000378:metabolism*; D019143:Evolution, Molecular; D015870:Gene Expression; D008958:Models, Molecular; D011123:Polyploidy; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D014157:Transcription Factors; D004247:DNA