Sensing actin dynamics: structural basis for G-actin-sensitive nuclear import of MAL

Biochem Biophys Res Commun. 2011 Oct 22;414(2):373-8. doi: 10.1016/j.bbrc.2011.09.079. Epub 2011 Sep 21.

Abstract

The coordination of cytoskeletal actin dynamics with gene expression reprogramming is emerging as a crucial mechanism to control diverse cellular processes, including cell migration, differentiation and neuronal circuit assembly. The actin-binding transcriptional coactivator MAL (also known as MRTF-A/MKL1/BSAC) senses G-actin concentration and transduces Rho GTPase signals to serum response factor (SRF). MAL rapidly shuttles between the cytoplasm and the nucleus in unstimulated cells but Rho-induced depletion of G-actin leads to MAL nuclear accumulation and activation of transcription of SRF:MAL-target genes. Although the molecular and structural basis of actin-regulated nucleocytoplasmic shuttling of MAL is not understood fully, it is proposed that nuclear import of MAL is mediated by importin α/β heterodimer, and that G-actin competes with importin α/β for the binding to MAL. Here we present structural, biochemical and cell biological evidence that MAL has a classical bipartite nuclear localization signal (NLS) in the N-terminal 'RPEL' domain containing Arg-Pro-X-X-X-Glu-Leu (RPEL) motifs. The NLS residues of MAL adopt an extended conformation and bind along the surface groove of importin-α, interacting with the major- and minor-NLS binding sites. We also present a crystal structure of wild-type MAL RPEL domain in complex with five G-actins. Comparison of the importin-α- and actin-complexes revealed that the binding of G-actins to MAL is associated with folding of NLS residues into a helical conformation that is inappropriate for importin-α recognition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / chemistry*
  • Actins / metabolism
  • Active Transport, Cell Nucleus
  • Animals
  • Cell Nucleus / metabolism*
  • Crystallography, X-Ray
  • Humans
  • Mice
  • Mutation
  • NIH 3T3 Cells
  • Nuclear Localization Signals / chemistry*
  • Nuclear Localization Signals / genetics
  • Nuclear Localization Signals / metabolism
  • Protein Structure, Tertiary
  • Rabbits
  • Trans-Activators / chemistry*
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • alpha Karyopherins / chemistry
  • alpha Karyopherins / metabolism

Substances

  • Actins
  • Mrtfa protein, mouse
  • Nuclear Localization Signals
  • Trans-Activators
  • alpha Karyopherins
  • karyopherin alpha 2