Chronic hepatitis, hepatocyte fragility, and increased soluble phosphoglycokeratins in transgenic mice expressing a keratin 18 conserved arginine mutant.
The two major intermediate filament proteins in glandular epithelia are keratin polypeptides 8 and 18 (K8/18). To evaluate the function and potential disease association of K18, we examined the effects of mutating a highly conserved arginine (arg89) of K18. Expression of K18 arg89-->his/cys and its normal K8 partner in cultured cells resulted in punctate staining as compared with the typical filaments obtained after expression of wild-type K8/18. Generation of transgenic mice expressing human K18 arg89-->cys resulted in marked disruption of liver and pancreas keratin filament networks. The most prominent histologic abnormalities were liver inflammation and necrosis that appeared at a young age in association with hepatocyte fragility and serum transaminase elevation. These effects were caused by the mutation since transgenic mice expressing wild-type human K18 showed a normal phenotype. A relative increase in the phosphorylation and glycosylation of detergent solubilized K8/18 was also noted in vitro and in transgenic animals that express mutant K18. Our results indicate that the highly conserved arg plays an important role in glandular keratin organization and tissue fragility as already described for epidermal keratins. Phosphorylation and glycosylation alterations in the arg mutant keratins may account for some of the potential changes in the cellular function of these proteins. Mice expressing mutant K18 provide a novel animal model for human chronic hepatitis, and for studying the tissue specific function(s) of K8/18.
Ku
N O
NO
Department of Medicine, VA Palo Alto Health Care System, California 94304, USA.
Michie
S
S
Oshima
R G
RG
Omary
M B
MB
D016475:3T3 Cells; D000818:Animals; D001120:Arginine / Q000502:physiology; D002460:Cell Line; D002908:Chronic Disease; D003545:Cysteine / Q000502:physiology; D003599:Cytoskeleton / Q000378:metabolism; D004195:Disease Models, Animal; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D019073:HT29 Cells; D006520:Hepatitis, Animal / Q000235:genetics*; D006639:Histidine / Q000502:physiology; D006801:Humans; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000502:physiology*; D007633:Keratins / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D012995:Solubility; D018411:Spodoptera
D006023:Glycoproteins; D007381:Intermediate Filament Proteins; D006639:Histidine; D007633:Keratins; D001120:Arginine; D003545:Cysteine