Disrupting microtubule network immobilizes amoeboid chemotactic receptor in the plasma membrane

S. De Keijzer, J. Galloway, G. S. Harms, Peter N Devreotes, Pablo A Iglesias

Research output: Contribution to journalArticle

Abstract

Signaling cascades are initiated in the plasma membrane via activation of one molecule by another. The interaction depends on the mutual availability of the molecules to each other and this is determined by their localization and lateral diffusion in the cell membrane. The cytoskeleton plays a very important role in this process by enhancing or restricting the possibility of the signaling partners to meet in the plasma membrane. In this study we explored the mode of diffusion of the cAMP receptor, cAR1, in the plasma membrane of Dictyostelium discoideum cells and how this is regulated by the cytoskeleton. Single-particle tracking of fluorescently labeled cAR1 using Total Internal Reflection Microscopy showed that 70% of the cAR1 molecules were mobile. These receptors showed directed motion and we demonstrate that this is not because of tracking along the actin cytoskeleton. Instead, destabilization of the microtubules abolished cAR1 mobility in the plasma membrane and this was confirmed by Fluorescence Recovery after Photobleaching. As a result of microtubule stabilization, one of the first downstream signaling events, the jump of the PH domain of CRAC, was decreased. These results suggest a role for microtubules in cAR1 dynamics and in the ability of cAR1 molecules to interact with their signaling partners.

Original languageEnglish (US)
Pages (from-to)1701-1708
Number of pages8
JournalBBA - Biomembranes
Volume1808
Issue number6
DOIs
StatePublished - Jun 2011

Fingerprint

Cell membranes
Microtubules
Cell Membrane
Molecules
Cytoskeleton
Fluorescence Recovery After Photobleaching
Photobleaching
Dictyostelium
Actin Cytoskeleton
Actins
Microscopy
Microscopic examination
Stabilization
Fluorescence
Chemical activation
Availability
Recovery

Keywords

  • Actin
  • Directed diffusion
  • FRAP
  • G-protein coupled receptor
  • Microtubules
  • Single-molecule TIRF microscopy

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Biophysics

Cite this

Disrupting microtubule network immobilizes amoeboid chemotactic receptor in the plasma membrane. / De Keijzer, S.; Galloway, J.; Harms, G. S.; Devreotes, Peter N; Iglesias, Pablo A.

In: BBA - Biomembranes, Vol. 1808, No. 6, 06.2011, p. 1701-1708.

Research output: Contribution to journalArticle

@article{b1c8c3eb509c449c947d31c99e6f9de9,
title = "Disrupting microtubule network immobilizes amoeboid chemotactic receptor in the plasma membrane",
abstract = "Signaling cascades are initiated in the plasma membrane via activation of one molecule by another. The interaction depends on the mutual availability of the molecules to each other and this is determined by their localization and lateral diffusion in the cell membrane. The cytoskeleton plays a very important role in this process by enhancing or restricting the possibility of the signaling partners to meet in the plasma membrane. In this study we explored the mode of diffusion of the cAMP receptor, cAR1, in the plasma membrane of Dictyostelium discoideum cells and how this is regulated by the cytoskeleton. Single-particle tracking of fluorescently labeled cAR1 using Total Internal Reflection Microscopy showed that 70{\%} of the cAR1 molecules were mobile. These receptors showed directed motion and we demonstrate that this is not because of tracking along the actin cytoskeleton. Instead, destabilization of the microtubules abolished cAR1 mobility in the plasma membrane and this was confirmed by Fluorescence Recovery after Photobleaching. As a result of microtubule stabilization, one of the first downstream signaling events, the jump of the PH domain of CRAC, was decreased. These results suggest a role for microtubules in cAR1 dynamics and in the ability of cAR1 molecules to interact with their signaling partners.",
keywords = "Actin, Directed diffusion, FRAP, G-protein coupled receptor, Microtubules, Single-molecule TIRF microscopy",
author = "{De Keijzer}, S. and J. Galloway and Harms, {G. S.} and Devreotes, {Peter N} and Iglesias, {Pablo A}",
year = "2011",
month = "6",
doi = "10.1016/j.bbamem.2011.02.009",
language = "English (US)",
volume = "1808",
pages = "1701--1708",
journal = "Biochimica et Biophysica Acta - Biomembranes",
issn = "0005-2736",
publisher = "Elsevier",
number = "6",

}

TY - JOUR

T1 - Disrupting microtubule network immobilizes amoeboid chemotactic receptor in the plasma membrane

AU - De Keijzer, S.

AU - Galloway, J.

AU - Harms, G. S.

AU - Devreotes, Peter N

AU - Iglesias, Pablo A

PY - 2011/6

Y1 - 2011/6

N2 - Signaling cascades are initiated in the plasma membrane via activation of one molecule by another. The interaction depends on the mutual availability of the molecules to each other and this is determined by their localization and lateral diffusion in the cell membrane. The cytoskeleton plays a very important role in this process by enhancing or restricting the possibility of the signaling partners to meet in the plasma membrane. In this study we explored the mode of diffusion of the cAMP receptor, cAR1, in the plasma membrane of Dictyostelium discoideum cells and how this is regulated by the cytoskeleton. Single-particle tracking of fluorescently labeled cAR1 using Total Internal Reflection Microscopy showed that 70% of the cAR1 molecules were mobile. These receptors showed directed motion and we demonstrate that this is not because of tracking along the actin cytoskeleton. Instead, destabilization of the microtubules abolished cAR1 mobility in the plasma membrane and this was confirmed by Fluorescence Recovery after Photobleaching. As a result of microtubule stabilization, one of the first downstream signaling events, the jump of the PH domain of CRAC, was decreased. These results suggest a role for microtubules in cAR1 dynamics and in the ability of cAR1 molecules to interact with their signaling partners.

AB - Signaling cascades are initiated in the plasma membrane via activation of one molecule by another. The interaction depends on the mutual availability of the molecules to each other and this is determined by their localization and lateral diffusion in the cell membrane. The cytoskeleton plays a very important role in this process by enhancing or restricting the possibility of the signaling partners to meet in the plasma membrane. In this study we explored the mode of diffusion of the cAMP receptor, cAR1, in the plasma membrane of Dictyostelium discoideum cells and how this is regulated by the cytoskeleton. Single-particle tracking of fluorescently labeled cAR1 using Total Internal Reflection Microscopy showed that 70% of the cAR1 molecules were mobile. These receptors showed directed motion and we demonstrate that this is not because of tracking along the actin cytoskeleton. Instead, destabilization of the microtubules abolished cAR1 mobility in the plasma membrane and this was confirmed by Fluorescence Recovery after Photobleaching. As a result of microtubule stabilization, one of the first downstream signaling events, the jump of the PH domain of CRAC, was decreased. These results suggest a role for microtubules in cAR1 dynamics and in the ability of cAR1 molecules to interact with their signaling partners.

KW - Actin

KW - Directed diffusion

KW - FRAP

KW - G-protein coupled receptor

KW - Microtubules

KW - Single-molecule TIRF microscopy

UR - http://www.scopus.com/inward/record.url?scp=79954926036&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79954926036&partnerID=8YFLogxK

U2 - 10.1016/j.bbamem.2011.02.009

DO - 10.1016/j.bbamem.2011.02.009

M3 - Article

C2 - 21334306

AN - SCOPUS:79954926036

VL - 1808

SP - 1701

EP - 1708

JO - Biochimica et Biophysica Acta - Biomembranes

JF - Biochimica et Biophysica Acta - Biomembranes

SN - 0005-2736

IS - 6

ER -