Am. J. Respir. Cell Mol. Biol., Vol 11, No. 1, Jul 1994, 1-6.
Dominant negative mutants: tools for the study of protein function in vitro and in vivo
D Sheppard
Lung Biology Center, University of California, San Francisco 94143-0854.
Powerful new approaches for the identification and sequencing of novel
cDNAs have produced a backlog of proteins seeking functions. Traditional
approaches for characterizing protein function (e.g., blocking monoclonal
antibodies and heterologous expression) have significant limitations,
especially in identifying the roles specific proteins play in vivo. An
alternative approach is to engineer mutations in the protein of interest
that abolish its function and that also inhibit the function of
simultaneously expressed wild-type protein (dominant negative mutations).
This approach has wide application to the study of a number of different
kinds of proteins but tends to be most effective for proteins that need to
assemble into multimers to be functional. Dominant negative mutants have
already provided insights into the molecular mechanisms of action of a
number of protein families, including hormone receptors, oncogenes, and
growth factor receptors, and have been identified as the cause of at least
a few autosomal dominant diseases. Expression of dominant negative mutants
under the control of highly active lung cell-specific promoters holds great
promise for the study of the roles specific proteins and protein families
play in lung development, health, and disease.
