What is it about?
Parafusin (PFUS) is involved in the membrane fusion machinery leading to exocytosis in Paramecium and was cloned and sequenced by us. PFUS is a member of phosphoglucomutase superfamily.PFUS coats the dense core of secretory vesicles (trichocysts) of Paramecium prior to exocytosis and may be isolated with organells.The membrane localization dissapears when exocytosis is stimulated in a calcium-dependent manner.
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Why is it important?
Molecular probes designed for the parafusin - the Paramecium exocytic-sensitive phosphoglycoprotein, gave distinct hybridization patterns in Saccharomyces cerevisiae genomic DNA when compared with different phosphoglucomutase specific probes. These include two probes identical to segments of yeast phosphoglucomutase (PGM) genes 1 and 2. Neither of the PGM probes revealed the 7.4 and 5.9 kb fragments in Bgl II-cut yeast DNA digest detected with the 1.6 kb cloned PFUS cDNA and oligonucleotide constructed to the PFUS region (insertion 3--I-3) not found in other species.
Perspectives
PCR amplification with PFUS-specific primers generated yeast DNA-species of the predicted molecular size which hybridized to the I-3 probe. A search of the yeast genome database produced an unassigned nucleotide sequence that showed 55% identity to parafusin gene and 37% identity to PGM2 (the major isoform of yeast phosphoglucomutase) within the amplified region.
Professor Elzbieta Wyroba
Nencki Institute of Experimental Biology
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This page is a summary of: A comparative hybridization analysis of yeast DNA with Paramecium parafusin- and different phosphoglucomutase-specific probes, Biochemistry and Cell Biology, January 2000, Canadian Science Publishing,
DOI: 10.1139/bcb-78-6-683.
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