3-Methyladenine inhibits autophagy in tobacco culture cells under sucrose starvation conditions

Chihiro Takatsuka, Yuko Inoue, Ken Matsuoka, Yuji Moriyasu

Research output: Contribution to journalArticlepeer-review

122 Citations (Scopus)


Tobacco (Nicotiana tabacum) culture cells perform autophagy and degrade cellular proteins in response to sucrose starvation. When protein degradation is blocked by the cysteine protease inhibitor E-64c, lysosomes containing particles of cytoplasm (autolysosomes) accumulate in the cells. Therefore, using light microscopy, we can determine whether cells have performed autophagy. In this study, we investigated whether or not 3-methyladenine (3-MA), which is a known inhibitor of autophagy in mammalian cells, blocks autophagy in tobacco culture cells. The accumulation of autolysosomes was blocked by the addition to the culture media of 5 mM 3-MA together with E-64c. We did not detect autolysosomes or structures thought to be involved with autophagy, such as autophagosomes, accumulating in these cells, as observed by electron microscopy. 3-MA blocked cellular protein degradation without any effect on cellular protease activity. In mammalian cells, phosphatidylinositol 3-kinase (PtdIns 3-kinase) is a putative target of 3-MA. The PtdIns 3-kinase inhibitors wortmannin and LY294002 also inhibited the accumulation of autolysosomes in tobacco culture cells. These results suggest that (1) 3-MA inhibits autophagy by blocking the formation of autophagosomes in tobacco culture cells, and (2) PtdIns 3-kinase is essential for autophagy in tobacco cells.

Original languageEnglish
Pages (from-to)265-274
Number of pages10
JournalPlant and Cell Physiology
Issue number3
Publication statusPublished - Mar 2004
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Physiology
  • Plant Science
  • Cell Biology


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