Large protein complexes retained in the ER are dislocated by non-COPII vesicles and degraded by selective autophagy

Multisubunit protein complexes are assembled in the endoplasmic reticulum (ER). Existing pools of single subunits and assembly intermediates ensure the efficient and rapid formation of complete complexes. While being kinetically beneficial, surplus components must be eliminated to prevent potentially harmful accumulation in the ER. Surplus single chains are cleared by the ubiquitin–proteasome system. However, the fate of not secreted assembly intermediates of multisubunit proteins remains elusive. Here we show by high-resolution double-label confocal immunofluorescence and immunogold electron microscopy that naturally occurring surplus fibrinogen Aα–γ assembly intermediates in HepG2 cells are dislocated together with EDEM1 from the ER to the cytoplasm in ER-derived vesicles not corresponding to COPII-coated vesicles originating from the transitional ER. This route corresponds to the novel ER exit path we have previously identified for EDEM1 (Zuber et al. Proc Natl Acad Sci USA 104:4407–4412, 2007). In the cytoplasm, detergent-insoluble aggregates of fibrinogen Aα–γ dimers develop that are targeted by the selective autophagy cargo receptors p62/SQSTM1 and NBR1. These aggregates are degraded by selective autophagy as directly demonstrated by high-resolution microscopy as well as biochemical analysis and inhibition of autophagy by siRNA and kinase inhibitors. Our findings demonstrate that different pathways exist in parallel for ER-to-cytoplasm dislocation and subsequent proteolytic degradation of large luminal protein complexes and of surplus luminal single-chain proteins. This implies that ER-associated protein degradation (ERAD) has a broader function in ER proteostasis and is not limited to the elimination of misfolded glycoproteins.     

Penicillamine induced changes in growing rats. II. liver parenchymal cell

Zusammenfassung d-Penicillamin als Kupferchelator bewirkt im Langzeitversuch eine isolierte Veränderung der stereologischen Parameter der Mitochondrien und der «microbodies». Zusammenhänge mit der Aktivitätshemmung kupferhaltiger Oxydasen in den beiden Zellorganellen werden vermutet.     

Genomic buffering mitigates the effects of deleterious mutations in bacteria

The relationship between the number of randomly accumulated mutations in a genome and fitness is a key parameter in evolutionary biology. Mutations may interact such that their combined effect on fitness is additive (no epistasis), reinforced (synergistic epistasis) or mitigated (antagonistic epistasis). We measured the decrease in fitness caused by increasing mutation number in the bacterium Salmonella typhimurium using a regulated, error-prone DNA polymerase (polymerase IV, DinB). As mutations accumulated, fitness costs increased at a diminishing rate. This suggests that random mutations interact such that their combined effect on fitness is mitigated and that the genome is buffered against the fitness reduction caused by accumulated mutations. Levels of the heat shock chaperones DnaK and GroEL increased in lineages that had accumulated many mutations, and experimental overproduction of GroEL further increased the fitness of lineages containing deleterious mutations. These findings suggest that overexpression of chaperones contributes to antagonistic epistasis.     

Einfluss der Ionenstärke auf das Immunpräzipitationsvermögen von Amphibien-Antikörpern

Summary Precipitating antibodies of amphibia are more dependent on the ionicity of the buffer system than the well known rabbit antibodies.     

Self-inhibiting extracellular proteins fromAspergillus oryzae

Zusammenfassung Mittels Ultrafiltration und Gelelektrophorese konnten 2 extrazellulare Protein-Fraktionen, Mol.-Gew. 27000–28000 daltons und 11000–12000 daltons, aus submersen Kulturen vonAspergillus oryzae gewonnen werden, welche Wachstumshemmung desselben Organismus bei Verabreichung zu Beginn der Kulturentwicklung verursachen

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Technical assistance by Mrs.U. Stangl and Mr.L. Berzaczy is greatly appreciated.

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These investigations are supported by the Austrian Fonds zur Förderung der wissenschaftlichen Forschung.     

Cerebral application of enkephalins

Summary Implantation of enkephalins A or B into the ventral thalamus or injection into the lateral ventricle of rats evoked only weak signs of stereotyped behavior, but did not cause gnawing.