Essential role of Drosophila black-pearl is mediated by its effects on mitochondrial respiration

Soumit Roy, Mary K. Short, E. Richard Stanley, Paul T. Jubinsky

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Black-pearl (Blp) is a highly conserved, essential inner-mitochondrial membrane protein. The yeast Blp homologue, Magmas/Pam16, is required for mitochondrial protein transport, growth, and survival. Our purpose was to determine the role of Drosophila Blp in mitochondrial function, cell survival, and proliferation. To this end, we performed mitotic recombination in Drosophila melanogaster, RNAi-mediated knockdown, MitoTracker staining, measurement of reactive oxygen species (ROS), flow cytometry, electron transport chain complex assays, and hemocyte isolation from Drosophila larvae. Proliferation-defective, Blp-deficient Drosophila Schneider cells exhibited mitochondrial membrane depolarization, a 60% decrease in ATP levels, increased amounts of ROS (3.5-fold), cell cycle arrest, and activation of autophagy that were associated with a selective 65% reduction of cytochrome c oxidase activity. N-acetyl cysteine (NAC) rescued Blp-RNAi-treated cells from cell cycle arrest, indicating that increased production of ROS is the primary cause of the proliferation and survival defects in Blp-depleted cells. blp hypomorph larvae had a 35% decreased number of plasmatocytes with a 45% reduced active mitochondrial staining and their viability was increased 2-fold by administration of NAC, which blocked melanotic lesions. Loss of Blp decreases cytochrome c oxidase activity and uncouples oxidative phosphorylation, causing ROS production, which selectively affects mitochondria-rich plasmatocyte survival and function, leading to melanotic lesions in Blp-deficient flies.

Original languageEnglish (US)
Pages (from-to)3822-3833
Number of pages12
JournalFASEB Journal
Volume26
Issue number9
DOIs
StatePublished - Sep 2012

Fingerprint

Drosophila
Reactive Oxygen Species
Respiration
Acetylcysteine
Cells
Mitochondrial Proteins
Mitochondrial Membranes
Electron Transport Complex IV
RNA Interference
Cell Cycle Checkpoints
Cysteine
Larva
Staining and Labeling
Hemocytes
Mitochondria
Flow cytometry
Oxidative Phosphorylation
Autophagy
Depolarization
Protein Transport

Keywords

  • Electron transport chain
  • Magmas
  • Melanotic inclusions
  • Pam 16
  • Reactive oxygen species

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Essential role of Drosophila black-pearl is mediated by its effects on mitochondrial respiration. / Roy, Soumit; Short, Mary K.; Stanley, E. Richard; Jubinsky, Paul T.

In: FASEB Journal, Vol. 26, No. 9, 09.2012, p. 3822-3833.

Research output: Contribution to journalArticle

Roy, Soumit ; Short, Mary K. ; Stanley, E. Richard ; Jubinsky, Paul T. / Essential role of Drosophila black-pearl is mediated by its effects on mitochondrial respiration. In: FASEB Journal. 2012 ; Vol. 26, No. 9. pp. 3822-3833.
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