Genetically encoded multimode reporter of adaptor complex 3 traffic in budding yeast

AP‐3 (adaptor complex 3) mediates traffic from the late Golgi or early endosomes to late endosomal compartments. In mammals, mutations in AP‐3 cause Hermansky‐Pudlak syndrome type 2, cyclic neutropenias, and a form of epileptic encephalopathy. In budding yeast, AP‐3 carries cargo directly from the trans‐Golgi to the lysosomal vacuole. Despite the pathway's importance and its discovery two decades ago, rapid screens and selections for AP‐3 mutants have not been available. We now report GNSI, a synthetic, genetically encoded reporter that allows rapid plate‐based assessment of AP‐3 functional deficiency, using either chromogenic or growth phenotype readouts. This system identifies defects in both the formation and consumption of AP‐3 carrier vesicles and is adaptable to high‐throughput screening or selection in both plate array and liquid batch culture formats. Episomal and integrating plasmids encoding GNSI have been submitted to the Addgene repository.

• Publication year

  2020

• Venue

  Traffic : the International Journal of Intracellular Transport

• Publication date

  2020-11-23

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1111/tra.12772PMID 33225520PMCID PMC8259613
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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