When virgin temperature-sensitive mutant cdc28 cells of the yeast Saccharomyces cerevisiae were incubated at restrictive temperatures, they continued to grow, reaching a maximum of 3.3-fold the original size after 24 h. The protein and RNA levels increased during the first 24 h, then gradually decreased. The cells that reached the maximal size lost proliferative activity and synthesized less protein. After a 72-h incubation, cellular components, protein, RNA and DNA, were progressively degraded, resulting in extensive fragmentation within 7 days. Light and electron microscopic observation revealed that cdc28 cells incubated at the restrictive temperature for 24 h were enriched with double-unit membranes in the cytoplasm, and the vacuoles were filled with autophagic body-like structures. After 7 days most cellular contents were lost, and the membrane systems were fragmented. The protein synthesis inhibitor cycloheximide added at 24 h inhibited degradation of protein for at least 7 days suggesting that protein synthesis was involved in the activation of autophagic death. All other temperature-sensitive cdc and secretory (sec) mutants tested showed similar morphological changes when arrested in the cell division cycle at the restrictive temperature. In contrast, the temperature-insensitive wild-type cells grew normally at 38 degrees C and only a few percent of them underwent autolysis 7 days after transfer to the stationary phase. These results suggested that the yeast cells underwent autophagic death when arrested at any stage of the cell division cycle, whereas those arrested at the stationary phase rarely underwent autophagic death.

• PMID: 8603680
• PubMed
• PubTator

Reference Type

Journal Article | Research Support, Non-U.S. Gov't

Authors

Motizuki M, Yokota S, Tsurugi K

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