Nitric oxide-dependent ribosomal RNA cleavage is associated with inhibition of ribosomal peptidyl transferase activity in ANA-1 murine macrophages.
NO can regulate specific cellular functions by altering transcriptional programs and protein reactivity. With respect to global cellular processes, NO has also been demonstrated to inhibit total protein synthesis and cell proliferation. The underlying mechanisms are unknown. In a system of ANA-1 murine macrophages, iNOS expression and NO production were induced by exposure to endotoxin (LPS). In selected instances, cells were exposed to an exogenous NO donor, S-nitroso-N-acetylpenicillamine or a substrate inhibitor of NO synthesis. Cellular exposure to NO, from both endogenous and exogenous sources, was associated with a significant time-dependent decrease in total protein synthesis and cell proliferation. Gene transcription was unaltered. In parallel with decreased protein synthesis, cells exhibited a distinctive cleavage pattern of 28S and 18S rRNA that were the result of two distinct cuts in both 28S and 18S rRNA. Total levels of intact 28S rRNA, 18S rRNA, and the composite 60S ribosome were significantly decreased in the setting of cell exposure to NO. Finally, 60S ribosome-associated peptidyl transferase activity, a key enzyme for peptide chain elongation, was also significantly decreased. Our data suggest that NO-mediated cleavage of 28S and 18S rRNA results in decreased 60S ribosome associated peptidyl transferase activity and inhibition of total protein synthesis.
Cai
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Department of Surgery, Georgetown University Medical Center, Washington, DC 20007, USA.
Guo
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Schroeder
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Punzalan
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Kuo
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D000818:Animals; D002455:Cell Division / Q000276:immunology; D002460:Cell Line; D004789:Enzyme Activation / Q000276:immunology; D006868:Hydrolysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology*; D010458:Peptidyl Transferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014176:Protein Biosynthesis; D011500:Protein Synthesis Inhibitors / Q000276:immunology; D011506:Proteins / Q000037:antagonists & inhibitors; D012335:RNA, Ribosomal / Q000276:immunology / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000096:biosynthesis / Q000378:metabolism*; D012270:Ribosomes / Q000201:enzymology; D014158:Transcription, Genetic / Q000276:immunology
D008070:Lipopolysaccharides; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; D009569:Nitric Oxide; D010458:Peptidyl Transferases