title: Molecular characterization and functional analysis of subunit 7 of eukaryotic initiation factor 3 from Eimeria tenella.
abstract: The initiation of translation in eukaryotic cells is stimulated by proteins known as initiation factors (eIFs). A structurally complex eIF composed of multiple subunits, eIF3 has been shown to have various functions in translation in a variety of eukaryotes. Until now, little is known about eIF3 in Eimeria tenella. Based on a previously identified expressed sequence tag(EST), we cloned the eIF3 subunit 7 gene (EteIF3s7) from E. tenella by rapid amplification of the cDNA ends(RACE). The 2278-bp full-length complementary DNA of EteIF3s7 contained a 1716-bp open reading frame (ORF) that encoded a 571-amino acid (aa) polypeptide. The EteIF3s7 protein contained the subunit 7 domain that is characteristic of members of the eIF3 zeta superfamily. The levels of EteIF3s7 messenger RNA and protein were higher in second generation merozoites than in sporulated oocysts, unsporulated oocysts, or sporozoites, and the EteIF3s7 protein was barely detectable in unsporulated oocysts. Our immunofluorescence analysis showed that the EteIF3s7 protein was uniformly distributed throughout the cytoplasm of sporozoites. After sporozoites were incubated in complete medium, the EteIF3s7 protein localized to the anterior region of the parasite. Following the first schizogenous division, the protein was uniformly dispersed in trophozoites, immature schizonts, and mature schizonts, and the EteIF3s7 protein was observed to be closely associated with the parasitophorous vacuole membrane. An anti-rEteIF3s7 polyclonal antibody inhibited the ability of E. tenella to invade DF-1 cells, which suggested that EteIF3s7 might be involved in host cell invasion and required for the growth of the parasite in the host.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Han
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Chunlin
authors.initials: C
authors.lastname: Kong
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Dong
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Shunhai
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Qiping
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhai
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Siting
authors.initials: S
authors.lastname: Liang
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Sha
authors.initials: S
authors.lastname: Li
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China.
authors.forename: Shihan
authors.initials: S
authors.lastname: Yang
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Minhang, Shanghai 200241, China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China. Electronic address: huangbing232@163.com.
authors.forename: Bing
authors.initials: B
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D001483:Base Sequence; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004306:Dose-Response Relationship, Immunologic; D016786:Eimeria tenella / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D039621:Eukaryotic Initiation Factor-3 / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D005347:Fibroblasts / Q000469:parasitology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification; D016053:RNA, Protozoan / Q000032:analysis / Q000302:isolation & purification; D011817:Rabbits; D013047:Specific Pathogen-Free Organisms
chemical_list: D000913:Antibodies, Protozoan; D018076:DNA, Complementary; D039621:Eukaryotic Initiation Factor-3; D012333:RNA, Messenger; D016053:RNA, Protozoan
keywords: Characterization; Eimeria tenella; Eukaryotic initiation factor 3; Invasion; Parasitophorous vacuole