Characterization of 14-3-3 isoforms expressed in the Echinococcus granulosus pathogenic larval stage.
The 14-3-3 protein family of eukaryotic regulators was studied in Echinococcus granulosus, the causative agent of cystic hydatid disease. These proteins mediate important cellular processes in eukaryotes and are expected to play important roles in parasite biology. Six isoforms of E. granulosus 14-3-3 genes and proteins (Eg14-3-3.1-6) were analyzed, and their phylogenetic relationships were established with bona fide 14-3-3 orthologous proteins from eukaryotic species. Eg14-3-3 isoforms with previous evidence of expression (Eg14-3-3.1-4) in E. granulosus pathogenic larval stage (metacestode) were cloned, and recombinant proteins were used for functional studies. These protein isoforms were detected in different components of E. granulosus metacestode, including interface components with the host. The roles that are played by Eg14-3-3 proteins in parasite biology were inferred from the repertoires of interacting proteins with each isoform, as assessed by gel overlay, cross-linking, and affinity chromatography assays. A total of 95 Eg14-3-3 protein ligands were identified by mass spectrometry. Eg14-3-3 isoforms have shared partners (44 proteins), indicating some overlapping functions; however, they also bind exclusive partners (51 proteins), suggesting Eg14-3-3 functional specialization. These ligand repertoires indicate the involvement of Eg14-3-3 proteins in multiple biochemical pathways in the E. granulosus metacestode and note some degree of isoform specialization.
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Aline
A
Teichmann
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Daiani M
DM
Vargas
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Karina M
KM
Monteiro
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Bruna V
BV
Meneghetti
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Cristine S
CS
Dutra
‡Escuela de Medicina Veterinaria, Facultad de Ecologia y Recursos Naturales, Universidad Andrés Bello, 8370251 Santiago, Chile.
Rodolfo
R
Paredes
§Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, 8389100 Santiago, Chile.
Norbel
N
Galanti
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Arnaldo
A
Zaha
†Laboratório de Genômica Estrutural e Funcional and Laboratório de Biologia Molecular de Cestódeos, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (UFRGS), 91501-970 Porto Alegre, RS, Brazil.
Henrique B
HB
Ferreira
D048948:14-3-3 Proteins / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001559:Benzhydryl Compounds; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D016000:Cluster Analysis; D004443:Echinococcosis / Q000469:parasitology*; D048209:Echinococcus granulosus / Q000378:metabolism* / Q000472:pathogenicity*; D005455:Fluorescent Antibody Technique; D040461:Gene Components; D005960:Glucosides; D015151:Immunoblotting; D007814:Larva / Q000378:metabolism / Q000472:pathogenicity; D008024:Ligands; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010802:Phylogeny; D025941:Protein Interaction Mapping; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism
D048948:14-3-3 Proteins; D001559:Benzhydryl Compounds; D005960:Glucosides; D008024:Ligands; D020033:Protein Isoforms; C570240:empagliflozin
14-3-3 proteins; Echinococcus granulosus; host−parasite interactions; protein−protein interactions