Mutation of a major keratin phosphorylation site predisposes to hepatotoxic injury in transgenic mice.
Simple epithelia express keratins 8 (K8) and 18 (K18) as their major intermediate filament (IF) proteins. One important physiologic function of K8/18 is to protect hepatocytes from drug-induced liver injury. Although the mechanism of this protection is unknown, marked K8/18 hyperphosphorylation occurs in association with a variety of cell stresses and during mitosis. This increase in keratin phosphorylation involves multiple sites including human K18 serine-(ser)52, which is a major K18 phosphorylation site. We studied the significance of keratin hyperphosphorylation and focused on K18 ser52 by generating transgenic mice that overexpress a human genomic K18 ser52--> ala mutant (S52A) and compared them with mice that overexpress, at similar levels, wild-type (WT) human K18. Abrogation of K18 ser52 phosphorylation did not affect filament organization after partial hepatectomy nor the ability of mouse livers to regenerate. However, exposure of S52A-expressing mice to the hepatotoxins, griseofulvin or microcystin, which are associated with K18 ser52 and other keratin phosphorylation changes, resulted in more dramatic hepatotoxicity as compared with WT K18-expressing mice. Our results demonstrate that K18 ser52 phosphorylation plays a physiologic role in protecting hepatocytes from stress-induced liver injury. Since hepatotoxins are associated with increased keratin phosphorylation at multiple sites, it is likely that unique sites aside from K18 ser52, and phosphorylation sites on other IF proteins, also participate in protection from cell stress.
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Department of Medicine, Veterans Administration Palo Alto Health Care System, Palo Alto, CA 94304, USA.
Michie
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SA
Soetikno
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RM
Resurreccion
E Z
EZ
Broome
R L
RL
Omary
M B
MB
D016475:3T3 Cells; D008841:Actin Cytoskeleton / Q000648:ultrastructure; D019943:Amino Acid Substitution; D000818:Animals; D056486:Chemical and Drug Induced Liver Injury / Q000235:genetics*; D020022:Genetic Predisposition to Disease; D006118:Griseofulvin / Q000633:toxicity; D006498:Hepatectomy; D006801:Humans; D007382:Intermediate Filaments / Q000502:physiology* / Q000648:ultrastructure; D007633:Keratins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008115:Liver Regeneration; D051379:Mice; D008822:Mice, Transgenic; D052998:Microcystins; D019319:Okadaic Acid / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000633:toxicity; D010766:Phosphorylation; D017354:Point Mutation; D011499:Protein Processing, Post-Translational
D052998:Microcystins; D010456:Peptides, Cyclic; D019319:Okadaic Acid; D006118:Griseofulvin; D007633:Keratins; C078588:microcystin