Nuclear localization of Formyl-Peptide Receptor 2 in human cancer cells

Fabio Cattaneo, Melania Parisi, Tiziana Fioretti, Daniela Sarnataro, Gabriella Esposito, Rosario Ammendola

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Current models of G protein-coupled receptors (GPCRs) signaling describe binding of external agonists to cell surface receptors which, in turn, trigger several biological responses. New paradigms indicate that GPCRs localize to and signal at the nucleus, thus regulating distinct signaling cascades. The formyl-peptide receptor FPR2 belongs to the GPCR super-family and is coupled to PTX-sensitive Gi proteins. We show by western blot analysis, immunofluorescence experiments and radioligand binding assays that FPR2 is expressed at nuclear level in CaLu-6 and AGS cells. Nuclear FPR2 is a functional receptor, since it participates in intra-nuclear signaling, as assessed by decreased G protein-FPR2 association and enhanced ERK2, c-Jun and c-Myc phosphorylation upon stimulation of intact nuclei with the FPR2 agonist, WKYMVm. We analyzed FPR2 sequence for the search of a nuclear localization sequence (NLS) and we found a stretch of basic aminoacids (227-KIHKK-231) in the third cytoplasmic loop of the receptor. We performed single (K230A) and multiple (H229A/K230A/K231A) mutagenesis of NLS. The constructs were individually overexpressed in HEK293 cells and immunofluorescence and western blot analysis showed that nuclear localization or translocation of FPR2 depends on the integrity of the H229 and K231 residues within the NLS.

Original languageEnglish
Pages (from-to)10-19
Number of pages10
JournalArchives of Biochemistry and Biophysics
Volume603
DOIs
Publication statusPublished - Aug 1 2016

Fingerprint

Formyl Peptide Receptor
G-Protein-Coupled Receptors
Cells
Fluorescent Antibody Technique
Western Blotting
Neoplasms
Radioligand Assay
Mutagenesis
Phosphorylation
HEK293 Cells
Cell Surface Receptors
Cytoplasmic and Nuclear Receptors
GTP-Binding Proteins
Assays
Association reactions
human FPR2 protein
Proteins
Experiments

Keywords

  • FPR2
  • G protein-coupled receptors
  • Nuclear localization
  • Signal transduction

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Nuclear localization of Formyl-Peptide Receptor 2 in human cancer cells. / Cattaneo, Fabio; Parisi, Melania; Fioretti, Tiziana; Sarnataro, Daniela; Esposito, Gabriella; Ammendola, Rosario.

In: Archives of Biochemistry and Biophysics, Vol. 603, 01.08.2016, p. 10-19.

Research output: Contribution to journalArticle

Cattaneo, Fabio ; Parisi, Melania ; Fioretti, Tiziana ; Sarnataro, Daniela ; Esposito, Gabriella ; Ammendola, Rosario. / Nuclear localization of Formyl-Peptide Receptor 2 in human cancer cells. In: Archives of Biochemistry and Biophysics. 2016 ; Vol. 603. pp. 10-19.
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