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Acinus integrates AKT1 and subapoptotic caspase activities to regulate basal autophagy.

Nandi N, Tyra LK, Stenesen D, Krämer H - J. Cell Biol. (2014)

Bottom Line: Acn stability also was regulated by AKT1-mediated phosphorylation.Physiologically, these flies showed improvements in processes known to be autophagy dependent, including increased starvation resistance, reduced Huntingtin-induced neurodegeneration, and prolonged life span.These data indicate that AKT1 and caspase-dependent regulation of Acn stability adjusts basal autophagy levels.

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Affiliation: Department of Neuroscience and Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390.

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Acn levels and activity is regulated by Dcp-1. (A–G) Micrographs of wild type (A) and eyes in which GMR-Gal4 drives expression of Acnwt (B), AcnD527A (C), Acnwt + Dcp-1–RNAi (D), Dcp-1-RNAi (E), Acnwt + Dredd-RNAi (F), Acnwt + Dronc-RNAi (G). Bar, 50 µm. (H–K′) Micrographs of larval eye discs stained for DNA and Acn (green) from OreR (H and H′), dcp-1 (I and I′), GMR-Gal4, UAS–Dcp-1–RNAi (J and J′), and GMR-Gal4, UAS-Dredd-1–RNAi (K and K′). (L–M″′) Micrographs of eye (L–L″′) and antennal (M–M″′) discs with clones of cells expressing Dcp-1–RNAi marked by RFP and stained for Acn and DNA. Arrow indicates the morphogenetic furrow and the broken line indicates the clone boundary. (N) Western blots of wild type (+/+) or dcp-1 larvae (−/−) expressing Myc-Acnwt or Myc-AcnD527A as indicated. Blots were probed for Myc or Actin. (O) Quantification of blots (n = 3) as shown in N. Bars: (H–M) 10 µm. Detailed genotypes and roughness quantification are given in Tables S1 and S2. Error bars show means ± SD. **, P < 0.01; ***, P < 0.001.
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fig4: Acn levels and activity is regulated by Dcp-1. (A–G) Micrographs of wild type (A) and eyes in which GMR-Gal4 drives expression of Acnwt (B), AcnD527A (C), Acnwt + Dcp-1–RNAi (D), Dcp-1-RNAi (E), Acnwt + Dredd-RNAi (F), Acnwt + Dronc-RNAi (G). Bar, 50 µm. (H–K′) Micrographs of larval eye discs stained for DNA and Acn (green) from OreR (H and H′), dcp-1 (I and I′), GMR-Gal4, UAS–Dcp-1–RNAi (J and J′), and GMR-Gal4, UAS-Dredd-1–RNAi (K and K′). (L–M″′) Micrographs of eye (L–L″′) and antennal (M–M″′) discs with clones of cells expressing Dcp-1–RNAi marked by RFP and stained for Acn and DNA. Arrow indicates the morphogenetic furrow and the broken line indicates the clone boundary. (N) Western blots of wild type (+/+) or dcp-1 larvae (−/−) expressing Myc-Acnwt or Myc-AcnD527A as indicated. Blots were probed for Myc or Actin. (O) Quantification of blots (n = 3) as shown in N. Bars: (H–M) 10 µm. Detailed genotypes and roughness quantification are given in Tables S1 and S2. Error bars show means ± SD. **, P < 0.01; ***, P < 0.001.

Mentions: To test which protease is responsible for cleaving Drosophila Acn, we performed a targeted RNAi screen. We took advantage of the rough eye phenotype resulting from eye-specific GMR-Gal4–driven expression of UAS-Acnwt at elevated temperature (28°C; Fig. 4, A and B). This phenotype was further enhanced when stabilized UAS-AcnD527A was expressed instead (Fig. 4 C). We reasoned that reduced activity of the protease responsible for Acn cleavage should enhance the roughness induced by UAS-Acnwt to a similar level to that of cleavage-resistant UAS-AcnD527A.


Acinus integrates AKT1 and subapoptotic caspase activities to regulate basal autophagy.

Nandi N, Tyra LK, Stenesen D, Krämer H - J. Cell Biol. (2014)

Acn levels and activity is regulated by Dcp-1. (A–G) Micrographs of wild type (A) and eyes in which GMR-Gal4 drives expression of Acnwt (B), AcnD527A (C), Acnwt + Dcp-1–RNAi (D), Dcp-1-RNAi (E), Acnwt + Dredd-RNAi (F), Acnwt + Dronc-RNAi (G). Bar, 50 µm. (H–K′) Micrographs of larval eye discs stained for DNA and Acn (green) from OreR (H and H′), dcp-1 (I and I′), GMR-Gal4, UAS–Dcp-1–RNAi (J and J′), and GMR-Gal4, UAS-Dredd-1–RNAi (K and K′). (L–M″′) Micrographs of eye (L–L″′) and antennal (M–M″′) discs with clones of cells expressing Dcp-1–RNAi marked by RFP and stained for Acn and DNA. Arrow indicates the morphogenetic furrow and the broken line indicates the clone boundary. (N) Western blots of wild type (+/+) or dcp-1 larvae (−/−) expressing Myc-Acnwt or Myc-AcnD527A as indicated. Blots were probed for Myc or Actin. (O) Quantification of blots (n = 3) as shown in N. Bars: (H–M) 10 µm. Detailed genotypes and roughness quantification are given in Tables S1 and S2. Error bars show means ± SD. **, P < 0.01; ***, P < 0.001.
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fig4: Acn levels and activity is regulated by Dcp-1. (A–G) Micrographs of wild type (A) and eyes in which GMR-Gal4 drives expression of Acnwt (B), AcnD527A (C), Acnwt + Dcp-1–RNAi (D), Dcp-1-RNAi (E), Acnwt + Dredd-RNAi (F), Acnwt + Dronc-RNAi (G). Bar, 50 µm. (H–K′) Micrographs of larval eye discs stained for DNA and Acn (green) from OreR (H and H′), dcp-1 (I and I′), GMR-Gal4, UAS–Dcp-1–RNAi (J and J′), and GMR-Gal4, UAS-Dredd-1–RNAi (K and K′). (L–M″′) Micrographs of eye (L–L″′) and antennal (M–M″′) discs with clones of cells expressing Dcp-1–RNAi marked by RFP and stained for Acn and DNA. Arrow indicates the morphogenetic furrow and the broken line indicates the clone boundary. (N) Western blots of wild type (+/+) or dcp-1 larvae (−/−) expressing Myc-Acnwt or Myc-AcnD527A as indicated. Blots were probed for Myc or Actin. (O) Quantification of blots (n = 3) as shown in N. Bars: (H–M) 10 µm. Detailed genotypes and roughness quantification are given in Tables S1 and S2. Error bars show means ± SD. **, P < 0.01; ***, P < 0.001.
Mentions: To test which protease is responsible for cleaving Drosophila Acn, we performed a targeted RNAi screen. We took advantage of the rough eye phenotype resulting from eye-specific GMR-Gal4–driven expression of UAS-Acnwt at elevated temperature (28°C; Fig. 4, A and B). This phenotype was further enhanced when stabilized UAS-AcnD527A was expressed instead (Fig. 4 C). We reasoned that reduced activity of the protease responsible for Acn cleavage should enhance the roughness induced by UAS-Acnwt to a similar level to that of cleavage-resistant UAS-AcnD527A.

Bottom Line: Acn stability also was regulated by AKT1-mediated phosphorylation.Physiologically, these flies showed improvements in processes known to be autophagy dependent, including increased starvation resistance, reduced Huntingtin-induced neurodegeneration, and prolonged life span.These data indicate that AKT1 and caspase-dependent regulation of Acn stability adjusts basal autophagy levels.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Neuroscience and Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390.

Show MeSH
Related in: MedlinePlus