A homolog of the E3 ubiquitin ligase Rbx1 is induced during hyperosmotic stress of salmon.
Juvenile salmon migrating from freshwater to the marine environment confront a marked change in environmental osmolality. Using differential display of mRNA expression, we cloned a 1.9-kb cDNA upregulated in isolated tissues of salmon exposed to the hyperosmotic stress associated with transition to the dehydrating marine environment. The cDNA codes for a 21-kDa protein, salmon hyperosmotic protein 21 (Shop21), with 98% identity to Rbx1, an E3 ubiquitin ligase; the protein also contains a novel 81-amino acid domain at the NH(2) terminus not found in Rbx1. Moderate hyperosmotic stress (24 h at 550 mosmol/kg) increased Shop21 transcript 10-fold in branchial lamellae, whereas no upregulation was observed under more severe stress (> or = 800 mosmol/kg). Expression of the gene also was observed in heart and kidney. Replacement of NaCl with mannitol, but not glycerol, also elicited an increase in Shop21 mRNA. Inhibition of the mitogen-activated protein kinase and mitogen-activated extracellular regulated kinase kinase signal transduction pathways failed to blunt the Shop21 response during hyperosmotic stress. Shop21 mRNA also accumulated during thermal stress but to a lesser extent than heat shock protein 70 mRNA. The potential importance of Shop21 to the living animal is suggested by marked upregulation of the gene in salmon after transfer to seawater. The results of these investigations suggest that Shop21 may have a role in targeting selected proteins (e.g., in freshwater ionocytes) nonessential for adaptation to seawater for removal via the proteasome pathway.
Pan
Feng
F
Department of Fisheries, Animal, and Veterinary Science, University of Rhode Island, Kingston, Rhode Island 02881, USA.
Zarate
Jacques
J
Bradley
Terence M
TM
D000222:Adaptation, Physiological / Q000187:drug effects / Q000502:physiology; D000818:Animals; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D005990:Glycerol / Q000494:pharmacology; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism*; D006358:Hot Temperature; D008025:Ligases / Q000235:genetics*; D008353:Mannitol / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D009994:Osmolar Concentration; D012333:RNA, Messenger / Q000378:metabolism; D012474:Salmon; D017386:Sequence Homology, Amino Acid; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D008353:Mannitol; D012965:Sodium Chloride; D044767:Ubiquitin-Protein Ligases; D008025:Ligases; D005990:Glycerol