Modulation of Cell Motility by Spatial Repositioning of Enzymatic ATP/ADP Exchange Capacity*
Remco van Horssen
1,
Edwin Janssen12,
Wilma Peters
,
Loes van de Pasch,
Mariska M. te Lindert,
Michiel M. T. van Dommelen,
Peter C. Linssen
,
Timo L. M. ten Hagen¶,
Jack A. M. Fransen, and
Bé Wieringa3
From the
Department of Cell Biology, Nijmegen Center for Molecular Life Sciences and Department of Hematology, Radboud University Nijmegen Medical Center, 6500 HB Nijmegen, The Netherlands and the ¶Department of Surgical Oncology, Erasmus MC, 3000 DR Rotterdam, The Netherlands
ATP is the "principal energy currency" in metabolism and the most versatile small molecular regulator of cellular activities. Although already much is known about the role of ATP in fundamental processes of living systems, data about its compartmentalization are rather scarce, and we still have only very limited understanding of whether patterns in the distribution of intracellular ATP concentration ("ATP inhomogeneity") do exist and have a regulatory role. Here we report on the analysis of coupling of local ATP supply to regulation of actomyosin behavior, a widespread and dynamic process with conspicuous high ATP dependence, which is central to cell shape changes and cell motility. As an experimental model, we use embryonic fibroblasts from knock-out mice without major ATP-ADP exchange enzymes, in which we (re)introduce the ATP/ADP exchange enzyme adenylate kinase-1 (AK1) and deliberately manipulate its spatial positioning by coupling to different artificial location tags. By transfection-complementation of AK1 variants and comparison with yellow fluorescent protein controls, we found that motility and spreading were enhanced in cells with AK1 with a focal contact guidance tag. Intermediary enhancement was observed in cells with membrane-targeted or cytosolic AK1. Use of a heterodimer-inducing approach for transient translocation of AK1 to focal contacts under conditions of constant global AK1 activity in the cell corroborated these results. Based on our findings with these model systems, we propose that local ATP supply in the cell periphery and "on site" fuelling of the actomyosin machinery, when maintained via enzymes involved in phosphoryl transfer, are codetermining factors in the control of cell motility.
Received for publication, September 9, 2008
, and in revised form, November 12, 2008.
This work is dedicated to the memory of Wilma Peters.
* This work was supported by Dutch Cancer Society Grant KUN 2002-1763. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. S1–S3 and Movies 1–9.
1 Both of these authors contributed equally to this work.
2 Present address: N.V. Organon, Schering-Plough Corp., Dept. of Target Discovery, PO Box 20, 5340 BH Oss, The Netherlands.
Deceased November 22, 2006.
3 To whom correspondence should be addressed: Dept. of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Medical Centre, PO Box 9101, 6500 HB Nijmegen, The Netherlands. Tel.: 31-24-3614329; Fax: 31-24-3615317; E-mail: b.wieringa{at}ncmls.ru.nl.
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