title: Modulation of cell motility by spatial repositioning of enzymatic ATP/ADP exchange capacity.
abstract: 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.
authors.affiliation: Department of Cell Biology, Nijmegen Center for Molecular Life Sciences and Department of Hematology, Radboud University Nijmegen Medical Center, 6500 HB Nijmegen, The Netherlands.
authors.forename: Remco
authors.initials: R
authors.lastname: van Horssen
authors.forename: Edwin
authors.initials: E
authors.lastname: Janssen
authors.forename: Wilma
authors.initials: W
authors.lastname: Peters
authors.forename: Loes
authors.initials: L
authors.lastname: van de Pasch
authors.forename: Mariska M Te
authors.initials: MM
authors.lastname: Lindert
authors.forename: Michiel M T
authors.initials: MM
authors.lastname: van Dommelen
authors.forename: Peter C
authors.initials: PC
authors.lastname: Linssen
authors.forename: Timo L M Ten
authors.initials: TL
authors.lastname: Hagen
authors.forename: Jack A M
authors.initials: JA
authors.lastname: Fransen
authors.forename: Bé
authors.initials: B
authors.lastname: Wieringa
mesh_terms: D000205:Actomyosin / Q000235:genetics / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000235:genetics / Q000378:metabolism*; D000263:Adenylate Kinase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000235:genetics; D002465:Cell Movement / Q000502:physiology*; D048430:Cell Shape / Q000502:physiology; D004622:Embryo, Mammalian / Q000166:cytology / Q000201:enzymology*; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D022001:Focal Adhesions / Q000201:enzymology / Q000235:genetics; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008954:Models, Biological; D021381:Protein Transport / Q000502:physiology
chemical_list: D007527:Isoenzymes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000205:Actomyosin; D000263:Adenylate Kinase; C110939:adenylate kinase 1