Actin filament attachments for sustained motility in vitro are maintained by filament bundling.

PloS One
Xiaohua Hu, Jeffrey R Kuhn

Abstract

We reconstructed cellular motility in vitro from individual proteins to investigate how actin filaments are organized at the leading edge. Using total internal reflection fluorescence microscopy of actin filaments, we tested how profilin, Arp2/3, and capping protein (CP) function together to propel thin glass nanofibers or beads coated with N-WASP WCA domains. Thin nanofibers produced wide comet tails that showed more structural variation in actin filament organization than did bead substrates. During sustained motility, physiological concentrations of Mg(2+) generated actin filament bundles that processively attached to the nanofiber. Reduction of total Mg(2+) abolished particle motility and actin attachment to the particle surface without affecting actin polymerization, Arp2/3 nucleation, or filament capping. Analysis of similar motility of microspheres showed that loss of filament bundling did not affect actin shell formation or symmetry breaking but eliminated sustained attachments between the comet tail and the particle surface. Addition of Mg(2+), Lys-Lys(2+), or fascin restored both comet tail attachment and sustained particle motility in low Mg(2+) buffers. TIRF microscopic analysis of filaments captured by WCA-coated b...Continue Reading

References

Oct 1, 1992·Archives of Biochemistry and Biophysics·A Romani, A Scarpa
Aug 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·G A DabiriF S Southwick
Jan 1, 1989·Cell Motility and the Cytoskeleton·D A KaiserT D Pollard
Sep 1, 1972·Experimental Cell Research·M Takeichi, T S Okada
Oct 1, 1982·The Journal of Cell Biology·T J RinkT Pozzan
Jul 3, 1995·Proceedings of the National Academy of Sciences of the United States of America·M B Goldberg, J A Theriot
Jan 1, 1993·Thrombosis Research·K MatsunoK Kobayashi
Apr 12, 1996·The Journal of Biological Chemistry·J X Tang, P A Janmey
Feb 1, 1997·Current Opinion in Cell Biology·S Higley, M Way
Nov 26, 1998·Experimental and Molecular Pathology·J TakayaY Kobayashi
Apr 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·L A CameronJ A Theriot
May 10, 2000·The Journal of Biological Chemistry·E F PlowJ W Smith
Aug 3, 2000·Frontiers in Bioscience : a Journal and Virtual Library·A M Romani, A Scarpa
Aug 23, 2000·The Journal of Cell Biology·J C Adams, M A Schwartz
Jan 9, 2001·Nature Cell Biology·J B MarchandH N Higgs
Mar 7, 2001·Current Biology : CB·L A CameronG G Borisy
Sep 5, 2001·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·D GoukassianV Andrés
Dec 14, 2001·Proceedings of the National Academy of Sciences of the United States of America·K J Amann, T D Pollard
Jan 25, 2002·Biophysical Journal·Richard B Dickinson, Daniel L Purich
Feb 5, 2002·Angiogenesis·B Murray, D J Wilson
May 17, 2002·Nature·Anne Bernheim-GroswasserCécile Sykes
Jan 29, 2003·The Journal of Cell Biology·Sebastian WiesnerDominique Pantaloni
Feb 5, 2003·The Journal of Cell Biology·Tatyana M SvitkinaGary G Borisy

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Citations

Nov 15, 2013·Molecular Biology of the Cell·Nimisha Khanduja, Jeffrey R Kuhn
Dec 5, 2014·Molecular Biology of the Cell·Meredith O SweeneyBruce L Goode
Jul 8, 2014·Current Opinion in Cell Biology·Brian R Graziano, Orion D Weiner
Nov 14, 2018·The Journal of Cell Biology·Xiaohua Hu, R Dyche Mullins

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Methods Mentioned

BETA
nucleotide exchange
motility assay
light scattering
GTPase
gel

Software Mentioned

MaxChelator
ImageJ
SlideBook

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