title: Protein synthesis inhibition induces cytoresistance in cultured human proximal tubular (HK-2) cells.
abstract: After sublethal injury, proximal tubular cells acquire resistance to further attack. This study evaluated whether this could be a possible consequence of decreased protein synthesis, a potential correlate of cell damage. To this end, cultured human proximal tubular cells (HK-2) were subjected to 0-24 h of protein synthesis inhibition (> 98%), either by adding protein synthesis inhibitors [cycloheximide (CH) or verrucarin A] or by inducing sublethal ATP depletion (antimycin A + 2-deoxyglucose). After 24 h of these treatments, significant resistance to Ca2+ ionophore/ATP depletion-induced attack was noted (assessed by vital dye exclusion, compared with normal cells). That < or = 6 h of protein synthesis inhibition caused no cytoresistance implied the importance of evolving protein depletion rather than nonspecific drug effects or protein synthesis inhibition per se. CH plus ATP depletion did not induce additive benefits, suggesting a common mechanism. Cytoresistance was dissociated from the extent of free Ca2+ loading and ATP depletion but was associated with a decrease in membrane deacylation. CH removal promptly restored protein synthesis and cytoresistance was lost; conversely, ATP recovery did not restore protein synthesis and cytoresistance persisted. The emergence of cytoresistance correlated with the disappearance/dephosphorylation of an unidentified 130-kDa tyrosine-phosphorylated protein/protein complex (denoted pp-130). The functional significance of this change was suggested by the fact that tyrosine phosphatase inhibition with orthovanadate maintained pp-130 expression and prevented the cytoresistant state. We conclude that protein synthesis inhibition in HK-2 cells can induce a cytoresistant state. Suppression in phospholipase activity and altered tyrosine phosphorylation events may have functional significance in this regard.
authors.affiliation: Fred Hutchinson Cancer Research Center, University of Washington, Seattle 98104, USA.
authors.forename: M
authors.initials: M
authors.lastname: Iwata
authors.forename: J
authors.initials: J
authors.lastname: Herrington
authors.forename: R A
authors.initials: RA
authors.lastname: Zager
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000968:Antimycin A / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects*; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D003847:Deoxyglucose / Q000494:pharmacology; D004347:Drug Interactions; D004988:Ethers, Cyclic / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D008970:Molecular Weight; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011506:Proteins / Q000302:isolation & purification; D014255:Trichothecenes / Q000494:pharmacology; D014638:Vanadates / Q000494:pharmacology
chemical_list: D004988:Ethers, Cyclic; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D014255:Trichothecenes; D019319:Okadaic Acid; D014638:Vanadates; D000968:Antimycin A; D000255:Adenosine Triphosphate; D003513:Cycloheximide; D003847:Deoxyglucose; D010749:Phosphoprotein Phosphatases; C005348:muconomycin A