Advanced

The phosphorylation motif at serine 225 governs the localization and function of sphingosine kinase 1 in resistance arteries

Lidington, Darcy; Peter, Bernhard Friedrich; Meissner, Anja LU ; Kroetsch, Jeffrey T.; Pitson, Stuart M.; Pohl, Ulrich and Bolz, Steffen Sebastian (2009) In Arteriosclerosis, Thrombosis, and Vascular Biology 29(11). p.1916-1922
Abstract

OBJECTIVE-: The purpose of this study was to characterize a phosphorylation motif at serine 225 as a molecular switch that regulates the pressure-dependent activation of sphingosine kinase 1 (Sk1) in resistance artery smooth muscle cells. METHODS AND RESULTS-: In isolated hamster gracilis muscle resistance arteries, pressure-dependent activation/translocation of Sk1 by ERK1/2 was critically dependent on its serine 225 phosphorylation site. Specifically, expression of Sk1s225A reduced resting and myogenic tone, resting Ca2+, pressure-induced Ca2+ elevations, and Ca 2+ sensitivity. The lack of function of the Sk1s225A mutant could not be entirely overcome by forced localization to the... (More)

OBJECTIVE-: The purpose of this study was to characterize a phosphorylation motif at serine 225 as a molecular switch that regulates the pressure-dependent activation of sphingosine kinase 1 (Sk1) in resistance artery smooth muscle cells. METHODS AND RESULTS-: In isolated hamster gracilis muscle resistance arteries, pressure-dependent activation/translocation of Sk1 by ERK1/2 was critically dependent on its serine 225 phosphorylation site. Specifically, expression of Sk1s225A reduced resting and myogenic tone, resting Ca2+, pressure-induced Ca2+ elevations, and Ca 2+ sensitivity. The lack of function of the Sk1s225A mutant could not be entirely overcome by forced localization to the plasma membrane via a myristoylation/palmitylation motif; the membrane anchor also significantly inhibited the function of the wild-type Sk1 enzyme. In both cases, Ca2+ sensitivity and myogenic tone were attenuated, whereas Ca 2+ handling was normalized/enhanced. These discrete effects are consistent with cell surface receptor-mediated effects (Ca2+ sensitivity) and intracellular effects of S1P (Ca2+ handling). Accordingly, S1P2 receptor inhibition (1μmol/L JTE013) attenuated myogenic tone without effect on Ca2+. CONCLUSIONS-: Translocation and precise subcellular positioning of Sk1 is essential for full Sk1 function; and two distinct S1P pools, proposed to be intra-and extracellular, contribute to the maintenance of vascular tone.

(Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
keywords
ERK1/2, Myogenic vasoconstriction, Signal transduction, Sphingosine-1-phosphate, Transfection
in
Arteriosclerosis, Thrombosis, and Vascular Biology
volume
29
issue
11
pages
7 pages
publisher
American Heart Association
external identifiers
  • scopus:73949131765
ISSN
1079-5642
DOI
10.1161/ATVBAHA.109.194803
language
English
LU publication?
no
id
49f8a710-7b34-4473-853b-3749c30a0e6b
date added to LUP
2017-05-23 22:25:48
date last changed
2017-07-30 05:25:36
@article{49f8a710-7b34-4473-853b-3749c30a0e6b,
  abstract     = {<p>OBJECTIVE-: The purpose of this study was to characterize a phosphorylation motif at serine 225 as a molecular switch that regulates the pressure-dependent activation of sphingosine kinase 1 (Sk1) in resistance artery smooth muscle cells. METHODS AND RESULTS-: In isolated hamster gracilis muscle resistance arteries, pressure-dependent activation/translocation of Sk1 by ERK1/2 was critically dependent on its serine 225 phosphorylation site. Specifically, expression of Sk1<sup>s225A</sup> reduced resting and myogenic tone, resting Ca<sup>2+</sup>, pressure-induced Ca<sup>2+</sup> elevations, and Ca <sup>2+</sup> sensitivity. The lack of function of the Sk1<sup>s225A</sup> mutant could not be entirely overcome by forced localization to the plasma membrane via a myristoylation/palmitylation motif; the membrane anchor also significantly inhibited the function of the wild-type Sk1 enzyme. In both cases, Ca<sup>2+</sup> sensitivity and myogenic tone were attenuated, whereas Ca <sup>2+</sup> handling was normalized/enhanced. These discrete effects are consistent with cell surface receptor-mediated effects (Ca<sup>2+</sup> sensitivity) and intracellular effects of S1P (Ca<sup>2+</sup> handling). Accordingly, S1P2 receptor inhibition (1μmol/L JTE013) attenuated myogenic tone without effect on Ca<sup>2+</sup>. CONCLUSIONS-: Translocation and precise subcellular positioning of Sk1 is essential for full Sk1 function; and two distinct S1P pools, proposed to be intra-and extracellular, contribute to the maintenance of vascular tone.</p>},
  author       = {Lidington, Darcy and Peter, Bernhard Friedrich and Meissner, Anja and Kroetsch, Jeffrey T. and Pitson, Stuart M. and Pohl, Ulrich and Bolz, Steffen Sebastian},
  issn         = {1079-5642},
  keyword      = {ERK1/2,Myogenic vasoconstriction,Signal transduction,Sphingosine-1-phosphate,Transfection},
  language     = {eng},
  number       = {11},
  pages        = {1916--1922},
  publisher    = {American Heart Association},
  series       = {Arteriosclerosis, Thrombosis, and Vascular Biology},
  title        = {The phosphorylation motif at serine 225 governs the localization and function of sphingosine kinase 1 in resistance arteries},
  url          = {http://dx.doi.org/10.1161/ATVBAHA.109.194803},
  volume       = {29},
  year         = {2009},
}