Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes

Vicki A. Sciorra, Simon A. Rudge, Jiyao Wang, Stuart McLaughlin, JoAnne Engebrecht, Andrew J. Morris

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Phospholipase D (PLD) generates lipid signals that coordinate membrane trafficking with cellular signaling. PLD activity in vitro and in vivo is dependent on phosphoinositides with a vicinal 4,5-phosphate pair. Yeast and mammalian PLDs contain an NH2-terminal pleckstrin homology (PH) domain that has been speculated to specify both subcellular localization and regulation of PLD activity through interaction with phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2). We report that mutation of the PH domains of yeast and mammalian PLD enzymes generates catalytically active PI(4,5)P2-regulated enzymes with impaired biological functions. Disruption of the PH domain of mammalian PLD2 results in relocalization of the protein from the PI(4,5)P2-containing plasma membrane to endosomes. As a result of this mislocalization, mutations within the PH domain render the protein unresponsive to activation in vivo. Furthermore, the integrity of the PH domain is vital for yeast PLD function in both meiosis and secretion. Binding of PLD2 to model membranes is enhanced by acidic phospholipids. Studies with PLD2-derived peptides suggest that this binding involves a previously identified polybasic motif that mediates activation of the enzyme by PI(4,5)P2. By comparison, the PLD2 PH domain binds PI(4,5)P2 with lower affinity but sufficient selectivity to function in concert with the polybasic motif to target the protein to PI(4,5)P2-rich membranes. Phosphoinositides therefore have a dual role in PLD regulation: membrane targeting mediated by the PH domain and stimulation of catalysis mediated by the polybasic motif.

Original languageEnglish (US)
Pages (from-to)1039-1049
Number of pages11
JournalJournal of Cell Biology
Volume159
Issue number6
DOIs
StatePublished - Dec 23 2002
Externally publishedYes

Fingerprint

4 alpha-glucanotransferase
Phospholipase D
Phosphatidylinositols
Yeasts
Membranes
Mutation
Proteins
Enzyme Activation
Endosomes
Meiosis
Pleckstrin Homology Domains
Catalysis
Phospholipids
Phosphates
Cell Membrane

Keywords

  • GTP-binding proteins
  • Membrane lipids
  • Phosphatidylinositol phosphates
  • Phospholipases
  • Signal transduction

ASJC Scopus subject areas

  • Cell Biology

Cite this

Sciorra, V. A., Rudge, S. A., Wang, J., McLaughlin, S., Engebrecht, J., & Morris, A. J. (2002). Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes. Journal of Cell Biology, 159(6), 1039-1049. https://doi.org/10.1083/jcb.200205056

Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes. / Sciorra, Vicki A.; Rudge, Simon A.; Wang, Jiyao; McLaughlin, Stuart; Engebrecht, JoAnne; Morris, Andrew J.

In: Journal of Cell Biology, Vol. 159, No. 6, 23.12.2002, p. 1039-1049.

Research output: Contribution to journalArticle

Sciorra, VA, Rudge, SA, Wang, J, McLaughlin, S, Engebrecht, J & Morris, AJ 2002, 'Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes', Journal of Cell Biology, vol. 159, no. 6, pp. 1039-1049. https://doi.org/10.1083/jcb.200205056
Sciorra VA, Rudge SA, Wang J, McLaughlin S, Engebrecht J, Morris AJ. Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes. Journal of Cell Biology. 2002 Dec 23;159(6):1039-1049. https://doi.org/10.1083/jcb.200205056
Sciorra, Vicki A. ; Rudge, Simon A. ; Wang, Jiyao ; McLaughlin, Stuart ; Engebrecht, JoAnne ; Morris, Andrew J. / Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes. In: Journal of Cell Biology. 2002 ; Vol. 159, No. 6. pp. 1039-1049.
@article{60674f9e0e0d42c0b039eaa94cc7aa7f,
title = "Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes",
abstract = "Phospholipase D (PLD) generates lipid signals that coordinate membrane trafficking with cellular signaling. PLD activity in vitro and in vivo is dependent on phosphoinositides with a vicinal 4,5-phosphate pair. Yeast and mammalian PLDs contain an NH2-terminal pleckstrin homology (PH) domain that has been speculated to specify both subcellular localization and regulation of PLD activity through interaction with phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2). We report that mutation of the PH domains of yeast and mammalian PLD enzymes generates catalytically active PI(4,5)P2-regulated enzymes with impaired biological functions. Disruption of the PH domain of mammalian PLD2 results in relocalization of the protein from the PI(4,5)P2-containing plasma membrane to endosomes. As a result of this mislocalization, mutations within the PH domain render the protein unresponsive to activation in vivo. Furthermore, the integrity of the PH domain is vital for yeast PLD function in both meiosis and secretion. Binding of PLD2 to model membranes is enhanced by acidic phospholipids. Studies with PLD2-derived peptides suggest that this binding involves a previously identified polybasic motif that mediates activation of the enzyme by PI(4,5)P2. By comparison, the PLD2 PH domain binds PI(4,5)P2 with lower affinity but sufficient selectivity to function in concert with the polybasic motif to target the protein to PI(4,5)P2-rich membranes. Phosphoinositides therefore have a dual role in PLD regulation: membrane targeting mediated by the PH domain and stimulation of catalysis mediated by the polybasic motif.",
keywords = "GTP-binding proteins, Membrane lipids, Phosphatidylinositol phosphates, Phospholipases, Signal transduction",
author = "Sciorra, {Vicki A.} and Rudge, {Simon A.} and Jiyao Wang and Stuart McLaughlin and JoAnne Engebrecht and Morris, {Andrew J.}",
year = "2002",
month = "12",
day = "23",
doi = "10.1083/jcb.200205056",
language = "English (US)",
volume = "159",
pages = "1039--1049",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "6",

}

TY - JOUR

T1 - Dual role for phosphoinositides in regulation of yeast and mammalian phospholipase D enzymes

AU - Sciorra, Vicki A.

AU - Rudge, Simon A.

AU - Wang, Jiyao

AU - McLaughlin, Stuart

AU - Engebrecht, JoAnne

AU - Morris, Andrew J.

PY - 2002/12/23

Y1 - 2002/12/23

N2 - Phospholipase D (PLD) generates lipid signals that coordinate membrane trafficking with cellular signaling. PLD activity in vitro and in vivo is dependent on phosphoinositides with a vicinal 4,5-phosphate pair. Yeast and mammalian PLDs contain an NH2-terminal pleckstrin homology (PH) domain that has been speculated to specify both subcellular localization and regulation of PLD activity through interaction with phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2). We report that mutation of the PH domains of yeast and mammalian PLD enzymes generates catalytically active PI(4,5)P2-regulated enzymes with impaired biological functions. Disruption of the PH domain of mammalian PLD2 results in relocalization of the protein from the PI(4,5)P2-containing plasma membrane to endosomes. As a result of this mislocalization, mutations within the PH domain render the protein unresponsive to activation in vivo. Furthermore, the integrity of the PH domain is vital for yeast PLD function in both meiosis and secretion. Binding of PLD2 to model membranes is enhanced by acidic phospholipids. Studies with PLD2-derived peptides suggest that this binding involves a previously identified polybasic motif that mediates activation of the enzyme by PI(4,5)P2. By comparison, the PLD2 PH domain binds PI(4,5)P2 with lower affinity but sufficient selectivity to function in concert with the polybasic motif to target the protein to PI(4,5)P2-rich membranes. Phosphoinositides therefore have a dual role in PLD regulation: membrane targeting mediated by the PH domain and stimulation of catalysis mediated by the polybasic motif.

AB - Phospholipase D (PLD) generates lipid signals that coordinate membrane trafficking with cellular signaling. PLD activity in vitro and in vivo is dependent on phosphoinositides with a vicinal 4,5-phosphate pair. Yeast and mammalian PLDs contain an NH2-terminal pleckstrin homology (PH) domain that has been speculated to specify both subcellular localization and regulation of PLD activity through interaction with phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2). We report that mutation of the PH domains of yeast and mammalian PLD enzymes generates catalytically active PI(4,5)P2-regulated enzymes with impaired biological functions. Disruption of the PH domain of mammalian PLD2 results in relocalization of the protein from the PI(4,5)P2-containing plasma membrane to endosomes. As a result of this mislocalization, mutations within the PH domain render the protein unresponsive to activation in vivo. Furthermore, the integrity of the PH domain is vital for yeast PLD function in both meiosis and secretion. Binding of PLD2 to model membranes is enhanced by acidic phospholipids. Studies with PLD2-derived peptides suggest that this binding involves a previously identified polybasic motif that mediates activation of the enzyme by PI(4,5)P2. By comparison, the PLD2 PH domain binds PI(4,5)P2 with lower affinity but sufficient selectivity to function in concert with the polybasic motif to target the protein to PI(4,5)P2-rich membranes. Phosphoinositides therefore have a dual role in PLD regulation: membrane targeting mediated by the PH domain and stimulation of catalysis mediated by the polybasic motif.

KW - GTP-binding proteins

KW - Membrane lipids

KW - Phosphatidylinositol phosphates

KW - Phospholipases

KW - Signal transduction

UR - http://www.scopus.com/inward/record.url?scp=0037164869&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037164869&partnerID=8YFLogxK

U2 - 10.1083/jcb.200205056

DO - 10.1083/jcb.200205056

M3 - Article

C2 - 12486109

AN - SCOPUS:0037164869

VL - 159

SP - 1039

EP - 1049

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 6

ER -