The motility-associated proteins GAP-43, MARCKS, and CAP-23 share unique targeting and surface activity-inducing properties

Exp Cell Res. 1997 Oct 10;236(1):103-16. doi: 10.1006/excr.1997.3709.

Abstract

Local regulation of the cortical cytoskeleton controls cell surface dynamics. GAP-43 and MARCKS are two abundant cytosolic protein kinase C substrates that are anchored to the cell membrane via acyl groups and interact with the cortical cytoskeleton. Each of them has been implicated in several forms of motility involving the cell surface. Although their primary sequences do not reveal significant homologies, GAP-43, MARCKS, and the cortical cytoskeleton-associated protein CAP-23 (in the following, the three proteins will be abbreviated as GMC) share a number of characteristic biochemical and biophysical properties and an unusual amino acid composition. In this study we determined whether GMC may be related functionally. In double-labeling immunocytochemistry experiments GMC accumulated at unique surface-associated structures, where they codistributed. In transfected cells GMC induced the same range of characteristic changes in cell morphology and cell surface activities, including prominent blebs and filopodia. These activities correlated with local accumulation of transgene and had characteristic features of locally elevated actin dynamics, including loss of stress fiber structures, accumulation of beta-(cytosolic) actin at cell surface protrusions, and dynamic blebbing activity. Analysis of appropriate deletion and fusion constructs revealed that the surface accumulation pattern and cell surface activities were correlated and that minimal structural requirements included acylation-mediated targeting to the cell membrane and the presence of a predominantly GMC-type sequence composition. Based on these experiments and on the results of previous studies on GAP-43, MARCKS, and CAP-23, we propose that GMC may define a class of functionally related proteins whose local accumulation promotes actin dynamics and the formation of dynamic structures at the cell periphery. Superimposed on these general properties, differences in the regulation of membrane association and binding properties of effector domains would confer individual properties to each of these proteins.

MeSH terms

  • 3T3 Cells / chemistry
  • 3T3 Cells / enzymology
  • 3T3 Cells / ultrastructure
  • Actins / metabolism
  • Animals
  • Antigens, Surface / analysis
  • Antigens, Surface / genetics
  • Antigens, Surface / metabolism*
  • COS Cells / chemistry
  • COS Cells / enzymology
  • COS Cells / ultrastructure
  • Calmodulin-Binding Proteins*
  • Cell Size
  • Chick Embryo
  • Cytoskeletal Proteins / analysis
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism*
  • Cytoskeleton / metabolism
  • GAP-43 Protein / analysis
  • GAP-43 Protein / genetics
  • GAP-43 Protein / metabolism*
  • Ganglia, Spinal / cytology
  • Gene Dosage
  • Intracellular Signaling Peptides and Proteins*
  • Membrane Proteins*
  • Mice
  • Microscopy, Electron
  • Myristoylated Alanine-Rich C Kinase Substrate
  • Nerve Tissue Proteins*
  • Protein Kinase C / analysis
  • Protein Kinase C / genetics
  • Protein Kinase C / metabolism
  • Proteins / analysis
  • Proteins / genetics
  • Proteins / metabolism*
  • Recombinant Proteins
  • Transfection

Substances

  • Actins
  • Antigens, Surface
  • Basp1 protein, mouse
  • Calmodulin-Binding Proteins
  • Cytoskeletal Proteins
  • GAP-43 Protein
  • Intracellular Signaling Peptides and Proteins
  • Marcks protein, mouse
  • Membrane Proteins
  • Nerve Tissue Proteins
  • Proteins
  • Recombinant Proteins
  • Myristoylated Alanine-Rich C Kinase Substrate
  • Protein Kinase C