Cycloheximide: Aspects of Inhibition of Protein Synthesis in Mammalian Cells

Ennis, Herbert L.; Lubin, Martin

doi:10.1126/science.146.3650.1474

Cited by 419 publications

(160 citation statements)

References 13 publications

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“…Cycloheximide, a eucaryote-specific inhibitor (Ennis and Lubin 1964), substantially inhibited the assimilation of [14C]bicarbonate, but not of thymidine or adenine; the assimilation rates of treated samples as percent of control samples were 9 1% for adenine, 88% for thymidine, and 38% for bicarbonate; in other words, < 10% of the adenine uptake was affected. The inhibition of bicarbonate assimilation suggests that most of the carbon fixation was by eucaryotes, in accordance with the observation that at this time of year, few if any cyanobacteria were present (L. Campbell pers.…”

Section: Resultsmentioning

confidence: 99%

Does adenine incorporation into nucleic acids measure total microbial production?1

Fuhrman

Ducklow

Kirchman

et al. 1986

Limnology & Oceanography

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Incorporation of radioactive adenine into DNA has recently been used as a measure of total microbial production in marine environments, sometimes with unexpected results. We have examined two of the assumptions on which the method is based, particularly regarding the distribution of adenine uptake and nucleic acid content in mixed natural microbial assemblages. Size fractionation, autoradiography, and metabolic inhibition data indicate that the requirement that bacteria, algae, and protozoa have a uniform uptake and metabolism of added dissolved adenine dots not appear to hold. In systems where the biomass and productivity were greatly dominated by large phytoplankton, bacteria took up almost all of the adenine added at nanomolar concentrations. A separate experiment with marine ciliates also showed adenine uptake by eucaryotes to be negligible compared to that by bacteria, The method requires a measurement of microbial adenosine triphosphate (ATP) specific activity. In our experiments this quantity would be dominated by ATP in the eucaryotes that took up negligible adenine. In this situation, mostly eucaryotic biomass and not production would influence the "total microbial production" measurement. Second, the C: DNA ratio of 50 used in reports to date is too high for natural bacteria, which have a ratio closer to 5. We conclude that the method is likely to have produced large overestimates of production and that variations (e.g. with depth) may not reflect growth rates, but rather a complicated combination of activities and ratios of eucaryotic : procaryotic biomasses.

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Section: Resultsmentioning

confidence: 99%

Does adenine incorporation into nucleic acids measure total microbial production?1

Fuhrman

Ducklow

Kirchman

et al. 1986

Limnology & Oceanography

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“…Like toxin, cycloheximide probably inhibits, reversibly, the action of transferase II in extracts from yeast as well as from mammalian cells. Like toxin, cycloheximide is completely without effect when added to the E. coli system (20). Neither toxin nor cycloheximide influence polysome breakdown or cause release of nascent peptide chains as is the case with puromycin (2,19).…”

Section: Effect Of Toxin Onmentioning

confidence: 97%

Studies on the Mode of Action of Diphtheria Toxin

Goor

Pappenheimer

Ames

1967

The Journal of Experimental Medicine

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In the preceding paper (1), we reported on the relative ability of various nucleotides related to nicofinamide adenine dinucleotide (NAD) to serve as cofactors for inhibition by diphtheria toxin of protein synthesis in cell-free extracts. Those few analogues which could replace NAD as activators of diphtheria toxin all proved to be nucleotides of demonstrated coenzyme activity. The results suggested that NAD and certain related substances are capable of interaction with the toxin protein. That diphtheria toxin does, in fact, reversibly bind one mole of NAD per mole of toxin was demonstrated by equilibrium dialysis and by gel filtration.In the present paper, we are reporting studies on the quantitative relationships between NAD, diphtheria toxin, and inhibition of peptide bond formation in cell-free extracts from various species. The data have led us to the conclusion that inhibition of protein synthesis, in vitro, results from reversible interaction between three components: toxin, NAD, and transferase II (2, 3). Reduction of NAD is not involved since it has been found that the inactivation of transferase II by toxin in mammalian cell extracts can be prevented and even reversed by relatively low concentrations of nicotinamide. Materials and MethodsReagents and Radioisotopes.--Materials used to determine amino acid incorporation in the cell-free systems were the same as in the preceding papers (1, 3). Nicotinamide, nicotinic acid, and pyridine-3-sulfonic acid were obtained from Nutritional Biochemical Corp., Cleveland,

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“…Cycloheximide is a protein synthesis inhibitor that inhibits the initiation of new peptide chains and the elongation of nascent peptides on ribosomes by different mechanisms (Ennis and Lubin, 1964).…”

Section: Lps Increases Nf-jb Activation Whereas Nf-jb Inhibition Attementioning

confidence: 99%

Bacterial endotoxin enhances colorectal cancer cell adhesion and invasion through TLR-4 and NF-κB-dependent activation of the urokinase plasminogen activator system

et al. 2009

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Perioperative exposure to lipopolysaccharide (LPS) is associated with accelerated metastatic colorectal tumour growth. LPS directly affects cells through Toll-like receptor 4 (TLR-4) and the transcription factor NF-kB. The urokinase plasminogen activator (u-PA) system is intimately implicated in tumour cell extracellular matrix (ECM) interactions fundamental to tumour progression. Thus we sought to determine if LPS directly induces accelerated tumour cell ECM adhesion and invasion through activation of the u-PA system and to elucidate the cellular pathways involved. Human colorectal tumour cell lines were stimulated with LPS. u-PA concentration, u-PA activity, active u-PA, surface urokinase plasminogen activator receptor (u-PAR) and TLR-4 expression were assessed by ELISA, colorimetric assay, western blot analysis and flow cytometry respectively. In vitro tumour cell vitronectin adhesion and ECM invasion were analysed by vitronectin adhesion assay and ECM invasion chambers. u-PA and u-PAR function was inhibited with anti u-PA antibodies or the selective u-PA inhibitors amiloride or WXC-340, TLR-4 by TLR-4-blocking antibodies and NF-kB by the selective NF-kB inhibitor SN-50. LPS upregulates u-PA and u-PAR in a dose-dependent manner, enhancing in vitro tumour cell vitronectin adhesion and ECM invasion by 440% (Po0.01). These effects were ameliorated by u-PA and u-PAR inhibition. LPS activates NF-kB through TLR-4. TLR-4 and NF-kB inhibition ameliorated LPS-enhanced u-PA and u-PAR expression, tumour cell vitronectin adhesion and ECM invasion. LPS promotes tumour cell ECM adhesion and invasion through activation of the u-PA system in a TLR-4-and NF-kB-dependent manner.

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Cycloheximide: Aspects of Inhibition of Protein Synthesis in Mammalian Cells

Cited by 419 publications

References 13 publications

Does adenine incorporation into nucleic acids measure total microbial production?1

Does adenine incorporation into nucleic acids measure total microbial production?1

Studies on the Mode of Action of Diphtheria Toxin

Bacterial endotoxin enhances colorectal cancer cell adhesion and invasion through TLR-4 and NF-κB-dependent activation of the urokinase plasminogen activator system

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