Identification of a phosphoinositide binding motif that mediates activation of mammalian and yeast phospholipase D isoenzymes

Vicki A. Sciorra, Simon A. Rudge, Glenn D. Prestwich, Michael A. Frohman, JoAnne Engebrecht, Andrew J. Morris

Research output: Contribution to journalArticle

131 Citations (Scopus)

Abstract

Phosphoinositides are both substrates for second messenger-generating enzymes and spatially localized membrane signals that mediate vital steps in signal transduction, cytoskeletal regulation and membrane trafficking. Phosphatidylcholine-specific phospholipase D (PLD) activity is stimulated by phosphoinositides, but the mechanism and physiological requirement for such stimulation to promote PLD-dependent cellular processes is not known. To address these issues, we have identified a site at which phosphoinositides interact with PLD and have assessed the role of this region in PLD function. This interacting motif contains critical basic amino acid residues that are required for stimulation of PLD activity by phosphoinositides. Although PLD alleles mutated at this site fail to bind to phosphoinositides in vitro, they are membrane-associated and properly localized within the cell but are inactive against cellular lipid substrates. Analogous mutations of this site in yeast PLD, Spo14p, result in enzymes that localize normally, but with catalytic activity that has dramatically reduced responsiveness to phosphoinositides. The level of responsiveness to phosphoinositides in vitro correlated with the ability of PLD to function in vivo. Taken together, these results provide the first evidence that phosphoinositide regulation of PLD activity observed in vitro is physiologically important in cellular processes in vivo including membrane trafficking and secretion.

Original languageEnglish (US)
Pages (from-to)5911-5921
Number of pages11
JournalEMBO Journal
Volume18
Issue number21
DOIs
StatePublished - Nov 1 1999
Externally publishedYes

Fingerprint

Phospholipase D
Phosphatidylinositols
Yeast
Isoenzymes
Yeasts
Chemical activation
Membranes
Basic Amino Acids
Signal transduction
Second Messenger Systems
Substrates
Enzymes
Phosphatidylcholines
Signal Transduction
Catalyst activity
Alleles
Lipids
Mutation

Keywords

  • Phosphatidylinositol 4,5-bisphosphate
  • Phospholipase D
  • Pleckstrin homology domain
  • Secretion

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Sciorra, V. A., Rudge, S. A., Prestwich, G. D., Frohman, M. A., Engebrecht, J., & Morris, A. J. (1999). Identification of a phosphoinositide binding motif that mediates activation of mammalian and yeast phospholipase D isoenzymes. EMBO Journal, 18(21), 5911-5921. https://doi.org/10.1093/emboj/18.21.5911

Identification of a phosphoinositide binding motif that mediates activation of mammalian and yeast phospholipase D isoenzymes. / Sciorra, Vicki A.; Rudge, Simon A.; Prestwich, Glenn D.; Frohman, Michael A.; Engebrecht, JoAnne; Morris, Andrew J.

In: EMBO Journal, Vol. 18, No. 21, 01.11.1999, p. 5911-5921.

Research output: Contribution to journalArticle

Sciorra, VA, Rudge, SA, Prestwich, GD, Frohman, MA, Engebrecht, J & Morris, AJ 1999, 'Identification of a phosphoinositide binding motif that mediates activation of mammalian and yeast phospholipase D isoenzymes', EMBO Journal, vol. 18, no. 21, pp. 5911-5921. https://doi.org/10.1093/emboj/18.21.5911
Sciorra, Vicki A. ; Rudge, Simon A. ; Prestwich, Glenn D. ; Frohman, Michael A. ; Engebrecht, JoAnne ; Morris, Andrew J. / Identification of a phosphoinositide binding motif that mediates activation of mammalian and yeast phospholipase D isoenzymes. In: EMBO Journal. 1999 ; Vol. 18, No. 21. pp. 5911-5921.
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