Gelsolin modulates phospholipase C activity in vivo through phospholipid binding

Hui Qiao Sun, Keng Mean Lin, Helen L. Yin

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

Gelsolin and CapG are actin regulatory proteins that remodel the cytoskeleton in response to phosphatidylinositol 4,5-bisphosphate (PIP2) and Ca2+ during agonist stimulation. A physiologically relevant rise in Ca2+ increases their affinity for PIP2 and can promote significant interactions with PIP2 in activated cells. This may impact divergent PIP2- dependent signaling processes at the level of substrate availability. We found that CapG overexpression enhances PDGF-stimulated phospholipase C(γ) (PLC(γ)) activity (Sun, H.-q., K. Kwiatkowska, D.C. Wooten, and H.L. Yin. 1995. J. Cell Biol. 129:147-156). In this paper, we examined the ability of gelsolin and CapG to compete with another PLC for PIP2 in live cells, in semi-intact cells, and in vitro. We found that CapG and gelsolin overexpression profoundly inhibited bradykinin-stimulated PLC(β). Inhibition occurred at or after the G protein activation step because overexpression also reduced the response to direct G protein activation with NaF. Bradykinin responsiveness was restored after cytosolic proteins, including gelsolin, leaked out of the overexpressing cells. Conversely, exogenous gelsolin added to permeabilized cells inhibited response in a dose-dependent manner. The washout and addback experiments clearly establish that excess gelsolin is the primary cause of PLC inhibition in cells. In vitro experiments showed that gelsolin and CapG stimulated as well as inhibited PLC(β), and only gelsolin domains containing PIP2-binding sites were effective. Inhibition was mitigated by increasing PIP2 concentration in a manner consistent with competition between gelsolin and PLC(β) for PIP2. Gelsolin and CapG also had biphasic effects on tyrosine kinase phosphorylated PLC(γ), although they inhibited PLC(γ) less than PLC(β). Our findings indicate that as PIP2 level and availability change during signaling, cross talk between PIP2-regulated proteins provides a selective mechanism for positive as well as negative regulation of the signal transduction cascade.

Original languageEnglish (US)
Pages (from-to)811-820
Number of pages10
JournalJournal of Cell Biology
Volume138
Issue number4
DOIs
StatePublished - Aug 25 1997

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Gelsolin
Type C Phospholipases
Phospholipids
Bradykinin
GTP-Binding Proteins
Phosphoinositide Phospholipase C
Solar System
Phosphatidylinositols
Cytoskeleton
Protein-Tyrosine Kinases
Actins
Signal Transduction
Proteins
Binding Sites

ASJC Scopus subject areas

  • Cell Biology

Cite this

Gelsolin modulates phospholipase C activity in vivo through phospholipid binding. / Sun, Hui Qiao; Lin, Keng Mean; Yin, Helen L.

In: Journal of Cell Biology, Vol. 138, No. 4, 25.08.1997, p. 811-820.

Research output: Contribution to journalArticle

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