Expression of a mutant ER-retained polytope membrane protein in cultured rat hepatocytes results in Mallory body formation

Abstract.

Mallory bodies represent cytokeratin-rich inclusion bodies which occur characteristically but not exclusively in human alcoholic liver disease and experimentally in mice during chronic intoxication with drugs. We report the first in vitro cell system of Mallory body induction. In clone 9 rat hepatocytes stably transfected to express an ER-retained T126M-aquaporin-2 (AQP2), on the mean 40% of the cells contained cytokeratin-rich inclusion bodies. By electron microscopy, their structure corresponded to that of genuine Mallory bodies. Such inclusion bodies were not detectable in clone 9 rat hepatocytes stably expressing a Golgi apparatus/lysosome-retained E258K-aquaporin-2. Proteasome inhibition increased the number of Mallory body-containing T126M-AQP2-expressing clone 9 hepatocytes to 60% on average. Proteasome inhibition in non-transfected, cytokeratin meshwork-forming clone 9 hepatocytes resulted in Mallory body formation on average in 6% of cells. Collectively, these data suggest that in the described in vitro cell system, Mallory body formation is induced by the presence of non-native protein conformers and point to the involvement of the proteasomal digestive system. The here reported in vitro system will be useful in studies about the biogenesis and progression of Mallory bodies, their relationship to aggresomes, and the role of inclusion bodies in the pathogenesis of cell damage.