A heterozygous mutation of beta-actin associated with neutrophil dysfunction and recurrent infection

Proc Natl Acad Sci U S A. 1999 Jul 20;96(15):8693-8. doi: 10.1073/pnas.96.15.8693.

Abstract

A human disorder caused by mutation in nonmuscle actin has not been reported. We report here a variant of nonmuscle actin in a female patient with recurrent infections, photosensitivity, and mental retardation. She also had abnormalities in neutrophil chemotaxis, superoxide production, and membrane potential response. Two-dimensional PAGE analysis of proteins from neutrophils and other cell types from this patient demonstrated a unique protein spot migrating at 42 kDa with pI shifted slightly to neutral relative to normal beta- and gamma-actin. Digestion peptide mapping and Western blotting showed this spot to be an abnormal actin. A full-length cDNA library was constructed by using mRNA from patient's cells and cDNA encoding the mutant beta-actin molecule was identified by an in vitro translation method. Sequencing of the clones demonstrated a G-1174 to A substitution, predicting a glutamic acid-364 to lysine substitution in beta-actin and eliminating a HinfI DNase restriction site found in normal beta-actin sequence. By HinfI digestion and by sequencing, the mutation in one allele of patient's genomic DNA was confirmed. Though no defect in cell-free polymerization of actin was detected, this defect lies in a domain important for binding to profilin and other actin-regulatory molecules. In fact, the mutant actin bound to profilin less efficiently than normal actin did. Heterozygous expression of mutant beta-actin in neutrophils and other cells of this patient may act in a dominant-negative fashion to adversely affect cellular activities dependent on the function of nonmuscle actin.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / chemistry
  • Actins / genetics*
  • Actins / metabolism
  • Chemotaxis / physiology
  • Child
  • Cloning, Molecular
  • Contractile Proteins*
  • DNA, Complementary / chemistry
  • Electrophoresis, Gel, Two-Dimensional
  • Female
  • Heterozygote
  • Humans
  • Infections / genetics*
  • Infections / immunology
  • Microfilament Proteins / metabolism
  • Mutation*
  • Neutrophils / chemistry
  • Neutrophils / physiology*
  • Polymorphism, Restriction Fragment Length
  • Profilins
  • Protein Binding
  • Protein Biosynthesis
  • Proteins / analysis
  • Sequence Analysis
  • Superoxides / metabolism

Substances

  • Actins
  • Contractile Proteins
  • DNA, Complementary
  • Microfilament Proteins
  • PFN1 protein, human
  • Profilins
  • Proteins
  • Superoxides