Plasma membrane phosphatidylinositol 4-phosphate and 4,5-bisphosphate determine the distribution and function of K-Ras4B but not H-Ras proteins

Gergo Gulyás, Glória Radvánszki, Rita Matuska, A. Balla, L. Hunyady, Tamas Balla, Péter Várnai

Research output: Contribution to journalArticle

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Abstract

Plasma membrane (PM) localization of Ras proteins is crucial for transmitting signals upon mitogen stimulation. Post-translational lipid modification of Ras proteins plays an important role in their recruitment to the PM. Electrostatic interactions between negatively charged PM phospholipids and basic amino acids found in K-Ras4B (K-Ras) but not in H-Ras are important for permanent K-Ras localization to the PM. Here, we investigated how acute depletion of negatively charged PM polyphosphoinositides (PPIns) from the PM alters the intracellular distribution and activity of K- and H-Ras proteins. PPIns depletion from the PM was achieved either by agonist-induced activation of phospholipase C β or with a rapamycin-inducible system in which various phosphatidylinositol phosphatases were recruited to the PM. Redistribution of the two Ras proteins was monitored with confocal microscopy or with a recently developed bioluminescence resonance energy transferbased approach involving fusion of the Ras C-terminal targeting sequences or the entire Ras proteins to Venus fluorescent protein. We found that PM PPIns depletion caused rapid translocation of K-Ras but not H-Ras from the PM to the Golgi. PM depletion of either phosphatidylinositol 4-phosphate (PtdIns4P) or PtdIns(4,5)P2 but not PtdIns(3,4,5)P3 was sufficient to evoke K-Ras translocation. This effect was diminished by deltarasin, an inhibitor of the Ras-phosphodiesterase interaction, or by simultaneous depletion of the Golgi PtdIns4P. The PPIns depletion decreased incorporation of [3H]leucine in K-Ras-expressing cells, suggesting that Golgi-localized K-Ras is not as signaling-competent as its PM-bound form. We conclude that PPIns in the PM are important regulators of K-Ras- mediated signals.

Original languageEnglish
Pages (from-to)18862-18877
Number of pages16
JournalJournal of Biological Chemistry
Volume292
Issue number46
DOIs
Publication statusPublished - Jan 1 2017

Fingerprint

ras Proteins
Cell membranes
Cell Membrane
Phosphatidylinositol Phosphates
phosphatidylinositol 4-phosphate
Venus
Phosphatidylinositol 4,5-Diphosphate
Bioluminescence
Basic Amino Acids
Phosphodiesterase Inhibitors
Confocal microscopy
Phosphoric Diester Hydrolases
Type C Phospholipases
Sirolimus
Post Translational Protein Processing
Phosphatidylinositols
Coulomb interactions
Static Electricity
Mitogens
Phosphoric Monoester Hydrolases

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Plasma membrane phosphatidylinositol 4-phosphate and 4,5-bisphosphate determine the distribution and function of K-Ras4B but not H-Ras proteins. / Gulyás, Gergo; Radvánszki, Glória; Matuska, Rita; Balla, A.; Hunyady, L.; Balla, Tamas; Várnai, Péter.

In: Journal of Biological Chemistry, Vol. 292, No. 46, 01.01.2017, p. 18862-18877.

Research output: Contribution to journalArticle

Gulyás, Gergo ; Radvánszki, Glória ; Matuska, Rita ; Balla, A. ; Hunyady, L. ; Balla, Tamas ; Várnai, Péter. / Plasma membrane phosphatidylinositol 4-phosphate and 4,5-bisphosphate determine the distribution and function of K-Ras4B but not H-Ras proteins. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 46. pp. 18862-18877.
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AU - Gulyás, Gergo

AU - Radvánszki, Glória

AU - Matuska, Rita

AU - Balla, A.

AU - Hunyady, L.

AU - Balla, Tamas

AU - Várnai, Péter

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