Energetic requirements for processive elongation of actin filaments by FH1FH2-formins

J Biol Chem. 2009 May 1;284(18):12533-40. doi: 10.1074/jbc.M808587200. Epub 2009 Feb 26.

Abstract

Formin-homology (FH) 2 domains from formin proteins associate processively with the barbed ends of actin filaments through many rounds of actin subunit addition before dissociating completely. Interaction of the actin monomer-binding protein profilin with the FH1 domain speeds processive barbed end elongation by FH2 domains. In this study, we examined the energetic requirements for fast processive elongation. In contrast to previous proposals, direct microscopic observations of single molecules of the formin Bni1p from Saccharomyces cerevisiae labeled with quantum dots showed that profilin is not required for formin-mediated processive elongation of growing barbed ends. ATP-actin subunits polymerized by Bni1p and profilin release the gamma-phosphate of ATP on average >2.5 min after becoming incorporated into filaments. Therefore, the release of gamma-phosphate from actin does not drive processive elongation. We compared experimentally observed rates of processive elongation by a number of different FH2 domains to kinetic computer simulations and found that actin subunit addition alone likely provides the energy for fast processive elongation of filaments mediated by FH1FH2-formin and profilin. We also studied the role of FH2 structure in processive elongation. We found that the flexible linker joining the two halves of the FH2 dimer has a strong influence on dissociation of formins from barbed ends but only a weak effect on elongation rates. Because formins are most vulnerable to dissociation during translocation along the growing barbed end, we propose that the flexible linker influences the lifetime of this translocative state.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Actin Cytoskeleton / genetics
  • Actin Cytoskeleton / metabolism*
  • Adenosine Triphosphate / genetics
  • Adenosine Triphosphate / metabolism*
  • Computer Simulation*
  • Energy Metabolism / physiology
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Models, Biological*
  • Profilins / genetics
  • Profilins / metabolism
  • Protein Structure, Tertiary / physiology
  • Quantum Dots
  • Saccharomyces cerevisiae / cytology
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • Bni1 protein, S cerevisiae
  • Microfilament Proteins
  • Profilins
  • Saccharomyces cerevisiae Proteins
  • Adenosine Triphosphate