Stage-specific expression of the 14-3-3 gene in Echinococcus multilocularis.
A cDNA expression library representing the metacestode developmental stage of the tapeworm Echinococcus multilocularis was immunoscreened with monospecific antibodies affinity purified following differential immunoblot analysis. Using this procedure, a metacestode-specific clone was isolated representing a 14-3-3 gene of the parasite, which is present as a single copy in the parasite genome. The identity of this clone was demonstrated by cross-reactivity of the recombinant E. multilocularis 14-3-3 protein with antibodies raised against a heterologous 14-3-3 protein from Saccharomyces cerevisiae. In addition, expression of the E. multilocularis 14-3-3 gene in the mutant S. cerevisiae strain, DS9-22, resulted in complementation of the phenotypic deficiency of this strain, thus demonstrating the functionality of the respective gene product. By reverse transcription-polymerase chain reaction (RT-PCR) we showed that the E. multilocularis 14-3-3 protein is about 10-fold overexpressed in the metacestode stage compared with the expression level in the adult stage. Immunolocalization of the 14-3-3 protein in E. multilocularis metacestodes revealed its predominant presence in the germinal layer of the parasite. The results of this study, taken together with the current knowledge on the 14-3-3 protein family, suggest that this parasite molecule may contribute to the promotion of the progressive, potentially unlimited growth behaviour of the E. multilocularis metacestode within the host tissue.
Siles-Lucas
M
M
Institute of Parasitology, University of Berne, Switzerland.
Felleisen
R S
RS
Hemphill
A
A
Wilson
W
W
Gottstein
B
B
D048948:14-3-3 Proteins; D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D003429:Cross Reactions; D004446:Echinococcus / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D017238:Genes, Helminth; D005816:Genetic Complementation Test; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D008018:Life Cycle Stages; D051379:Mice; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development; D014446:Tyrosine 3-Monooxygenase
D048948:14-3-3 Proteins; D000947:Antigens, Helminth; D015801:Helminth Proteins; D011506:Proteins; D014446:Tyrosine 3-Monooxygenase