Cycloheximide Is Not a Specific Inhibitor of Protein Synthesis <i>in Vivo</i>

McMahon, Daniel

doi:10.1104/pp.55.5.815

Cited by 116 publications

(51 citation statements)

References 35 publications

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“…In the presence of cycloheximide, both ACC accumulation and C2H4 production from exogenous ACC were inhibited. This could suggest that continuous protein synthesis is required for the operation of this pathway; however, this inhibitor is known to have other side effects (10). Taken together, the data presented here indicate that the pathway for C2H4 production in chilled cucumbers is the same as has been proposed for a number of higher plant tissues (1, 4, 9, 19).…”

Section: Resultssupporting

confidence: 67%

Ethylene Production by Chilled Cucumbers (Cucumis sativus L.)

Wang¹,

Adams²

1980

Plant Physiol.

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Chilling at 2.5 C accelerated the synthesis of 1-aminocyclopropane-lcarboxylic acid (ACC) and C2H. production in cucumber fruit. Skin tissue contained higher levels of ACC and was more sensitive to chilling than was cortex tissue. Accumulation of ACC in chilled tissue was detected after 1 day of chilling and remained elevated even after C2H4 production started to decline. These data suggest that ACC synthesis is readily stimulated by chilling, whereas the system that converts ACC to C2H4 is vulnerable to chilling injury. Chilling-induced C2H4 production was inhibited by aminoethoxyvinylglycine, sodium benzoate, propyl gallate, 2,4dinitrophenol, carbonyl cyanide m-chlorophenylhydrazone, and cycloheximide. The utilization of methionine for ACC formation and chilling-induced C2H1 biosynthesis was established using L-13,4-14Clmethionine. Chilled tissue had a higher capacity to convert L-13,4-14Clmethionine to ACC and C2H4 than did nonchilled tissue.Chilling temperatures have been observed to stimulate the production of C2H4 by tissues of a number of plants, including citrus, avocados, and pears (6, 13, 15). However, very little is known about the mechanism of this chilling-induced C2H4 production.It has recently been recognized that ACC2 is a precursor for C2H4 in many higher plant tissues (1,4,9). This unusual amino acid is produced from methionine via SAM. The proposed sequence for the pathway of C2H4 biosynthesis thus becomes: methionine -* SAM --ACC -* C2H4. The study presented here was initiated to determine if this pathway accounts for the production of C2H4 from chilled tissue and to identify the step(s) that are enhanced by chilling temperature.The C2H4 biosynthetic pathway is inhibited by AVG (7), uncouplers of oxidative phosphorylation (12), and free radical scavengers (2, 3). Therefore, the effect of these inhibitors on C2H4 production and ACC levels in chilled cucumbers has been examined. samples were divided into two lots and stored at 2.5 C or 13 C. MATERIALSEach day, development of chilling injury symptoms was evaluated in five cucumbers from each treatment group that were transferred to a room maintained at 20 C. Chilling injury was evaluated as previously described (14), and the severity of injury was rated on a scale of 1 to 5 (1, no abnormality; 2, trace; 3, slight; 4, moderate; and 5, severe injury).C2H4 and ACC Determination. C2H4 production by intact cucumbers stored at 2.5 C or 13 C was measured daily. Duplicate samples of two cucumbers each from each temperature treatment were transferred to a room maintained at 25 C and were allowed to warm to room temperature. Then they were placed in 1-liter chambers and sealed for 1 h. Gas samples then were taken for C2H4 determination by GC (11). After C2H4 determination, 3 g skin and 3 g cortex tissue were taken from each cucumber and assayed for ACC content by the method of Lizada and Yang (8).Incorporation of L-13,4-14CIMethionine. Three g skin tissue were placed in a 50-ml Erlenmeyer flask with 4 ml 10 mm Mes buffer (pH 6.0) containing 2% suc...

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Section: Resultssupporting

confidence: 67%

Ethylene Production by Chilled Cucumbers (Cucumis sativus L.)

Wang¹,

Adams²

1980

Plant Physiol.

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“…~974;Marcus & Sekellick, 1975). In addition, cycloheximide might affect intracellular processes other than inhibition of protein synthesis (McMahon, 1975).…”

Section: Introductionmentioning

confidence: 99%

Interferon Action III. The Rate of Primary Transcription of Vesicular Stomatitis Virus is Inhibited by Interferon Action

Marcus

Sekellick

1978

Journal of General Virology

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SUMMARYTranscription of vesicular stomatitis virus (VSV) in Vero cells was confined to the synthesis of parentally-derived mRNA (primary transcription) by the use of cycloheximide and/or a ts mutant, G4I(IV), at a non-permissive temperature (40 °C). More transcripts accumulated in the presence of cycloheximide than in its absence. This so-called 'cycloheximide effect' results from higher rates of virus transcription sustained for longer periods of time. The rate of VSV transcription initially increases linearly for I to 2 h after infection. Interferon reduces this rate (~_ fourfold with 50 units/ml interferon) irrespective of the presence or absence of cycloheximide. The VSV mRNA transcripts synthesized in mock-or interferontreated cells were equal in size and had an equivalent half-life of 17 h at 40 °C. It seems likely that once transcription is initiated in interferon-treated cells, it is completed successfully.Since interferon reduces the rate of early VSV primary transcript synthesis to below that achieved in the presence of cycloheximide, we conclude that interferon has an effect on transcription beyond that attributable solely to protein synthesis inhibition. We postulate that interferon decreases the probability of initiating virus transcription. Virus mRNA escaping this facet of interferon action may then encounter other facets such as post-transcriptional modification and/or inhibition of translation. However, the mandatory sequence of primary transcription-~ primary translation for negative-strand viruses like VSV dictates that the overall inhibitory effect of interferon on translation would derive in part from this prior inhibition of transcription. Thus, to apply the term 'primary effect' to one particular facet of interferon action may not always be meaningful.

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“…In order to clarify the timing of synthesis of protein molecules which are necessary for the induction of DNA synthesis in potato discs, CH was applied to the discs at variously delayed time after slicing and its effect on the rate of DNA synthesis was determined. Specificity of the action of CH has been questioned by some investigators (16,19), but since inhibitors with other action mechanisms, e.g. puromycin, pfluorophenylalanine, canavanine, and ethionine, also blocked the induction of DNA synthesis in potato discs as previously shown (22), it is very likely that CH acts on DNA synthesis through inhibiting protein synthesis.…”

Section: Discussionmentioning

confidence: 99%

Induction of Deoxyribonucleic Acid Synthesis in Potato Tuber Slices

Watanabe¹,

Imaseki²

1976

Plant Physiol.

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Timing of protein synthesis which is a prerequisite to DNA synthesis induced in potato tuber tissue (Solanum tuberosum L.) by cut injury has been studied using cycloheximide. The induction of DNA synthesis which was measured by incorporation of 3H-thymidine was completely inhibited when the inhibitor was applied to the tuber discs immediately after slicing. When the application of cycloheximide was delayed for 6 hours or more after slcing, DNA synthesis was observed but its rate was reduced to 20% of control. The inhibitory effect of cycloheximide, however, rapidly decreased when the inhibitor was applied at 6 or less hours immediately prior to determination of DNA synthesis. (1,10). RNA and protein syntheses were reported to be involved in the development of wound respiration in potato tuber slices and the necessary protein was synthesized in a very

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Cycloheximide Is Not a Specific Inhibitor of Protein Synthesis in Vivo

Cited by 116 publications

References 35 publications

Ethylene Production by Chilled Cucumbers (Cucumis sativus L.)

Ethylene Production by Chilled Cucumbers (Cucumis sativus L.)

Interferon Action III. The Rate of Primary Transcription of Vesicular Stomatitis Virus is Inhibited by Interferon Action

Induction of Deoxyribonucleic Acid Synthesis in Potato Tuber Slices

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