A Systematic RNAi Screen Reveals a Novel Role of a Spindle Assembly Checkpoint Protein BuGZ in Synaptic Transmission in C. elegans

Synaptic vesicles (SV) store various neurotransmitters that are released at the synapse. The molecular mechanisms of biogenesis, exocytosis, and endocytosis for SV, however, remain largely elusive. In this study, using Complex Object Parametric Analysis and Sorter (COPAS) to monitor the fluorescence of synapto-pHluorin (SpH), we performed a whole-genome RNAi screen in C. elegans to identify novel genetic modulators in SV cycling. One hundred seventy six genes that up-regulating SpH fluorescence and 96 genes that down-regulating SpH fluorescence were identified after multi-round screen. Among these genes, B0035.1 (bugz-1) encodes ortholog of mammalian C2H2 zinc-finger protein BuGZ/ZNF207, which is a spindle assembly checkpoint protein essential for mitosis in human cells. Combining electrophysiology, imaging and behavioral assays, we reveal that depletion of BuGZ-1 results in defects in locomotion. We further demonstrate that BuGZ-1 promotes SV recycling by regulating the expression levels of endocytosis-related genes such as rab11.1. Therefore, we have identified a bunch of potential genetic modulators in SV cycling, and revealed an unexpected role of BuGZ-1 in regulating synaptic transmission.

• Publication year

  2017

• Venue

  Frontiers in Molecular Neuroscience

• Publication date

  2017-05-11

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.3389/fnmol.2017.00141PMID 28553202PMCID 5425591
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• BuGZ-1 promotes synaptic vesicle recycling by regulating expression of endocytosis-related genes including rab11.1.

  Confidence 0.93

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• BuGZ-1 depletion causes locomotion defects in C. elegans.

  Confidence 0.95

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• A COPAS-based whole-genome RNAi screen using synapto-pHluorin identified 176 genes that increased fluorescence and 96 genes that decreased fluorescence, implicating them in synaptic vesicle cycling.

  Confidence 0.97

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review

• bugz-1

  gene, protein

  A C. elegans gene/protein referenced as the BuGZ/ZNF207 ortholog in the abstract.

  Aliases: B0035.1, BuGZ, ZNF207

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• copas

  instrumentation, method

  The Complex Object Parametric Analysis and Sorter platform used here to measure fluorescence in C. elegans samples.

  Aliases: Complex Object Parametric Analysis and Sorter

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• locomotion defects

  phenotype, behavioral phenotype

  An impairment in worm movement used as a behavioral phenotype in the experiments.

  Aliases: defects in locomotion

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• rab11.1

  gene

  A C. elegans gene mentioned as an endocytosis-related expression target in the abstract.

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• synaptic vesicle cycling

  biological process

  The sequence of synaptic vesicle exocytosis, endocytosis, and reuse at neuronal synapses.

  Aliases: SV cycling

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• synaptic vesicle recycling

  biological process

  The reuse of synaptic vesicles after endocytosis to sustain neurotransmission.

  Aliases: SV recycling

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• synapto-phluorin

  reporter, molecular tool

  A pH-sensitive fluorescent reporter used to monitor synaptic vesicle exocytosis and recycling.

  Aliases: SpH

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review
• whole-genome rnai screen

  method

  A genome-wide RNA interference screening approach used to perturb gene function one gene at a time.

  Aliases: genome-wide RNAi screen

  박진우 (dztg5apj7m) extraction뀨 (7c402c1b98) reviewKiller Whale (322360f1c1) reviewB (s683577b42) reviewq (76h6bfydm6) review

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