Relative importance of βcyto-and γcyto-actin in primary mouse embryonic fibroblasts

Xiaobai Patrinostro, Allison R. O'Rourke, Christopher M Chamberlain, Branden S Moriarity, Benjamin J. Perrin, James M Ervasti

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

The highly homologous β (βcyto) and γ (γcyto) cytoplasmic actins are hypothesized to carry out both redundant and unique essential functions, but studies using targeted gene knockout and siRNA-mediated transcript knockdown to examine βcyto-and γcyto-isoform-specific functions in various cell types have yielded conflicting data. Here we quantitatively characterized actin transcript and protein levels, as well as cellular phenotypes, in both geneand transcript-targeted primary mouse embryonic fibroblasts. We found that the smooth muscle αsm-actin isoform was the dominantly expressed actin isoform in WT primary fibroblasts and was also the most dramatically up-regulated in primary βcyto-or β/γcyto-actin double-knockout fibroblasts. Gene targeting of βcyto-actin, but not γcyto-actin, led to greatly decreased cell proliferation, decreased levels of cellular ATP, and increased serum response factor signaling in primary fibroblasts, whereas immortalization induced by SV40 large T antigen supported fibroblast proliferation in the absence of βcyto-actin. Consistent with in vivo gene-targeting studies in mice, both gene-and transcript-targeting approaches demonstrate that the loss of βcyto-actin protein is more disruptive to primary fibroblast function than is the loss of γcyto-actin.

Original languageEnglish (US)
Pages (from-to)771-782
Number of pages12
JournalMolecular biology of the cell
Volume28
Issue number6
DOIs
StatePublished - Mar 15 2017

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