Background
Mutations in the human gene encoding the neuron-specific Eag1 (KV10.1; KCNH1) potassium channel are linked to congenital neurodevelopmental diseases. Disease-causing mutant Eag1 channels manifest aberrant gating function and defective protein homeostasis. Both the E3 ubiquitin ligase cullin 7 (Cul7) and the small acid protein 14-3-3 serve as binding partners of Eag1. Cul7 mediates proteasomal and lysosomal degradation of Eag1 protein, whereas over-expression of 14-3-3 notably reduces Eag1 channel activity. It remains unclear whether 14-3-3 may also contribute to Eag1 protein homeostasis.
Results
In human cell line and native rat neurons, disruptions of endogenous 14-3-3 function with the peptide inhibitor difopein or specific RNA interference up-regulated Eag1 protein level in a transcription-independent manner. Difopein hindered Eag1 protein ubiquitination at the endoplasmic reticulum and the plasma membrane, effectively promoting the stability of both immature and mature Eag1 proteins. Suppression of endogenous 14-3-3 function also reduced excitotoxicity-associated Eag1 degradation in neurons. Difopein diminished Cul7-mediated Eag1 degradation, and Cul7 knock-down abolished the effect of difopein on Eag1. Inhibition of endogenous 14-3-3 function substantially perturbed the interaction of Eag1 with Cul7. Further structural analyses suggested that the intracellular Per-Arnt-Sim (PAS) domain and cyclic nucleotide-binding homology domain (CNBHD) of Eag1 are essential for the regulatory effect of 14-3-3 proteins. Significantly, suppression of endogenous 14-3-3 function reduced Cul7-mediated degradation of disease-associated Eag1 mutant proteins.
Conclusion
Overall these results highlight a chaperone-like role of endogenous 14-3-3 proteins in regulating Eag1 protein homeostasis, as well as a therapeutic potential of 14-3-3 modulators in correcting defective protein expression of disease-causing Eag1 mutants.
The aim of the present study was to investigate the effect of a mixture of dietary chicken essence and fish oil on possible lowering of blood and hepatic lipids in hamsters. Forty male hamsters were randomly divided into four groups and fed a chow diet, high fat/cholesterol diet (HFC), HFC diets containing 4% (HFC4) or 8% (HFC8) fish oil with essence of chicken for 42 days. The experimental diets contained 10% (w/w} fat. Hamsters fed HFC diets containing either 4 or 8% fish oil with essence of chicken showed significantly decreased serum triacylglycerols (TG} compared to hamsters fed HFC diets. In addition, hamsters fed HFC8 diets exhibited significantly reduced hepatic TG than those fed HFC diet. However, hamsters fed HFC4 or HFC8 diets significantly increased low density lipoprotein cholesterol (LDL-C) and decreased high density lipoprotein cholesterol (HDL-C) when compared with those on HFC diet. These results suggest that 4% (or more} fish oil with essence of chicken might significantly lower serum TG and hepatic TG in hamsters, but seem to increase serum cholesterol levels on HFC diets.
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