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Molecular cloning of the plc1 + gene of Schizosaccharomyces pombe , which encodes a putative phosphoinositide‐specific phospholipase C
Author(s) -
Andoh Tomoko,
YokoO Takehiko,
Matsui Yasushi,
TohE Akio
Publication year - 1995
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320110209
Subject(s) - schizosaccharomyces pombe , biology , gene , open reading frame , schizosaccharomyces , cloning (programming) , molecular cloning , peptide sequence , genbank , yeast , phospholipase c , saccharomyces cerevisiae , biochemistry , microbiology and biotechnology , enzyme , computer science , programming language
Exploiting the polymerase chain reaction, we have isolated a gene that encodes a putative phosphoinositide‐specific phospholipase C (PLC) of the fission yeast Schizosaccharomyces pombe . Inspection of the nucleotide sequence of the gene revealed an open reading frame that can encode a polypeptide of 899 amino acid residues with a calculated molecular mass of 102 kDa. This putative polypeptide contains both the X and Y regions that are conserved among three classes of mammalian PLC, and also contains a presumptive Ca 2+ ‐binding site (an E‐F hand motif). The structure of the putative protein is most similar to that of the δ class of PLC isozymes. To investigate the role of this gene, designated plc1 + , gene disruption was carried out by interrupting the coding region with the ura4 + marker. Growth of plc1 cells was temperature‐sensitive in rich medium, and cells could not grow in synthetic medium. Expression of the PLC1 gene of Saccharomyces cerevisiae suppressed the growth defect phenotype of plc1 − cells, a strong suggestion that the plc1 + gene encodes PLC. The PLC1 sequence appears in the public data libraries, DDBJ GenBank, EMBL under the following Accession Number: D38309.