Characterization of a plasma membrane-associated phosphoinositide-specific phospholipase C from soybean

Plant J. 1995 Sep;8(3):381-90. doi: 10.1046/j.1365-313x.1995.08030381.x.

Abstract

Phosphoinositide-specific phospholipase C (PI-PLC) is a key signal transducing enzyme which generates the second messengers inositol trisphosphate and diacylglycerol in mammalian cells. A cDNA clone (PI-PLC1) encoding a phosphoinositide-specific phospholipase C was isolated from soybean by screening a cDNA expression library using an anti-(plasma membrane) serum. Genomic DNA gel blot analysis suggested that the corresponding gene is a member of a multigene family. The deduced amino acid sequence of the soybean PI-PLC1 isozyme contains the conserved X and Y regions, found in other PI-PLCs. It is closely related to mammalian delta-type PI-PLCs, Dictyostelium discoideum PI-PLC and yeast PI-PLC1 in terms of the arrangement of the conserved region. Unlike mammalian delta-type PI-PLCs and yeast PI-PLC1, the putative Ca(2+)-binding site of the soybean PI-PLC1 is located in the region spanning the X and Y domains, and the N-terminal region is truncated. FLAG epitope-tagged PI-PLC1 fusion protein purified from transgenic tobacco plants showed phosphoinositide-specific phospholipase C activity. Heterologous expression of the soybean PI-PLC1 cDNA in a yeast PI-PLC1 deletion mutant complemented the lethality phenotype of haploid PI-PLC1 disruptants. Immunoblot analysis of the cell fractions prepared from transgenic tobacco plants over-expressing the FLAG epitope-tagged PI-PLC1 fusion protein indicated that the protein encoded by the PI-PLC1 cDNA was localized in the cytosol and plasma membrane.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Membrane / enzymology
  • DNA, Complementary
  • Gene Library
  • Genes, Plant
  • Isoenzymes / biosynthesis
  • Isoenzymes / chemistry
  • Isoenzymes / isolation & purification
  • Mammals
  • Molecular Sequence Data
  • Multigene Family
  • Phosphatidylinositol Diacylglycerol-Lyase
  • Phosphoinositide Phospholipase C
  • Phosphoric Diester Hydrolases / biosynthesis*
  • Phosphoric Diester Hydrolases / chemistry*
  • Phosphoric Diester Hydrolases / isolation & purification
  • Plants, Genetically Modified
  • Plants, Toxic
  • Plasmids
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / chemistry
  • Saccharomyces cerevisiae
  • Second Messenger Systems
  • Signal Transduction
  • Soybeans / enzymology*
  • Soybeans / genetics
  • Tobacco / enzymology

Substances

  • DNA, Complementary
  • Isoenzymes
  • Recombinant Fusion Proteins
  • Phosphoric Diester Hydrolases
  • Phosphoinositide Phospholipase C
  • Phosphatidylinositol Diacylglycerol-Lyase

Associated data

  • GENBANK/U25027