Cloning and molecular characterization of a cDNA clone coding for Trichomonas vaginalis alpha-actinin and intracellular localization of the protein.
We have identified and sequenced a cDNA clone coding for Trichomonas vaginalis alpha-actinin. Analysis of the obtained sequence revealed that the 2,857-nucleotide-long cDNA contained an open reading frame encoding 849 amino acids which showed consistent homology with alpha-actinins of different species. Such homology was particularly significant in regions which have been reported to represent the actin-binding and Ca2+-binding domains in other alpha-actinins. The deduced protein was also characterized by the presence of a divergent central region thought to play a role in its high immunogenicity. A study of protein localization performed by immunofluorescence revealed that the protein is diffusely distributed throughout the T. vaginalis cytoplasm when the cell is pear shaped. When parasites adhere and transform into the amoeboid morphology, the protein is located only in areas close to the cytoplasmic membrane and colocalizes with actin. Concomitantly with transformation into the amoeboid morphology, alpha-actinin mRNA expression is upregulated.
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Division of Experimental and Clinical Microbiology, Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
Rappelli
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Delogu
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Carta
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Cappuccinelli
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Fiori
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D000185:Actinin / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D002451:Cell Compartmentation; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000648:ultrastructure; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D014246:Trichomonas vaginalis / Q000235:genetics* / Q000648:ultrastructure
D018076:DNA, Complementary; D000939:Epitopes; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D000185:Actinin