Cycloheximide is a naturally occurring fungicide produced by Streptomyces griseus that inhibits eukaryotic protein synthesis by blocking translational elongation through interference with tRNA and mRNA movement at the ribosome. Widely used in in vitro biomedical research due to its rapid and reversible effects, it is unsuitable for human therapeutic use because of toxic side effects including DNA damage, teratogenesis, and reproductive toxicity such as birth defects. Although previously applied as a fungicide in agriculture, its use has declined over health concerns. Cycloheximide decontamination can be performed with soapy water or sodium bicarbonate. It is classified as an extremely hazardous substance under the U.S. Emergency Planning and Community Right-to-Know Act.
Discovery
Cycloheximide was reported in 1946 by Alma Joslyn Whiffen-Barksdale at the Upjohn Company.4
Experimental applications
Cycloheximide can be used as an experimental tool in molecular biology to determine the half-life of a protein. Treating cells with cycloheximide in a time-course experiment followed by western blotting of the cell lysates for the protein of interest can show differences in protein half-life. Cycloheximide treatment provides the ability to observe the half-life of a protein without confounding contributions from transcription or translation. Irreversible analogues of cycloheximide have also been reported.5
Mitochondrial protein synthesis is resistant to inhibition by cycloheximide. On the other hand chloramphenicol inhibits mitochondrial (and bacterial) protein synthesis, but synthesis on cytoplasmic ribosomes is resistant. Before genomes were available, these inhibitors were used to determine which mitochondrial proteins were synthesized in the mitochondria from mitochondrial genes.67
Cycloheximide is used as a plant growth regulator to stimulate ethylene production. It is used as a rodenticide and other animal pesticide. It is also used in media to detect unwanted bacteria in beer fermentation by suppressing yeasts and molds growth in test medium.
The translational elongation freezing properties of cycloheximide are also used for ribosome profiling / translational profiling. Translation is halted via the addition of cycloheximide, and the DNA/RNA in the cell is then nuclease treated. The ribosome-bound parts of RNA can then be sequenced.
Cycloheximide has also been used to make isolation of bacteria from environmental samples easier.8
Spectrum of fungal susceptibility
Cycloheximide has been used to isolate dermatophytes and inhibit the growth of fungi in brewing test media. The following represents susceptibility data for a few commonly targeted fungi:9
- Candida albicans: 12.5 μg/ml
- Mycosphaerella graminicola: 47.2 μg/ml – 85.4 μg/ml
- Saccharomyces cerevisiae: 0.05 μg/ml – 1.6 μg/ml
- Neoscytalidium dimidiatum is an Athlete's foot like infection resistant to most antifungals but is rather sensitive to cycloheximide, so, it should be cultured in a medium free of cycloheximide.
See also
References
Müller, Franz; Ackermann, Peter; Margot, Paul (2012). "Fungicides, Agricultural, 2. Individual Fungicides". Ullmann's Encyclopedia of Industrial Chemistry. Weinheim: Wiley-VCH. doi:10.1002/14356007.o12_o06. ISBN 978-3527306732. 978-3527306732 ↩
"TOXNET". toxnet.nlm.nih.gov. Archived from the original on 2007-05-22. Retrieved 2007-05-03. https://web.archive.org/web/20070522110734/http://toxnet.nlm.nih.gov/index.html ↩
"40 C.F.R.: Appendix A to Part 355—The List of Extremely Hazardous Substances and Their Threshold Planning Quantities" (PDF). Code of Federal Regulations (July 1, 2008 ed.). Government Printing Office. Archived from the original (PDF) on February 25, 2012. Retrieved October 29, 2011. https://web.archive.org/web/20120225051612/http://edocket.access.gpo.gov/cfr_2008/julqtr/pdf/40cfr355AppA.pdf ↩
New York Botanical Gardens. "Alma Whiffen Barksdale Records (RG5)". nybg.org. Retrieved 1 March 2017. http://www.nybg.org/library/finding_guide/archv/barksdale_rg5f.html ↩
Park, Yongho; Koga, Yumi; Su, Cindy; Waterbury, Amanda L.; Johnny, Christopher L.; Liau, Brian B. (2019-04-08). "Versatile Synthetic Route to Cycloheximide and Analogues That Potently Inhibit Translation Elongation". Angewandte Chemie International Edition. 58 (16): 5387–5391. doi:10.1002/anie.201901386. ISSN 1433-7851. PMID 30802354. https://onlinelibrary.wiley.com/doi/10.1002/anie.201901386 ↩
Weiss H, Sebald W, and Bücher T (1971). "Cycloheximide resistant incorporation of amino acids into a polypeptide of the cytochrome oxidase of Neurospora crassa" (PDF). Eur. J. Biochem. 22 (1): 19–26. doi:10.1111/j.1432-1033.1971.tb01509.x. PMID 4329217. https://opus.bibliothek.uni-wuerzburg.de/files/5835/Sebald68.pdf ↩
Sebald W, Weiss H, and Jackl G (1972). "Inhibition of the Assembly of Cytochrome Oxidase in Neurospora crassa by Chloramphenicol". Eur. J. Biochem. 30 (3): 413–417. doi:10.1111/j.1432-1033.1972.tb02112.x. PMID 4344826. https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-62852 ↩
Sands, D. C.; Rovira, A. D. (September 1970). "Isolation of Fluorescent Pseudomonads with a Selective Medium". Applied Microbiology. 20 (3): 513–514. doi:10.1128/am.20.3.513-514.1970. ISSN 0003-6919. PMC 376970. PMID 16349887. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC376970 ↩
"Cycloheximide – The Antimicrobial Index Knowledgebase – TOKU-E". antibiotics.toku-e.com. http://antibiotics.toku-e.com/antimicrobial_548.html ↩