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

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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 β- and γ-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 β-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 β-actin and eliminating a HinfI DNase restriction site found in normal β-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 β-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.