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

Area of Science:

• Cell Biology
• Molecular Biology
• Genetics

Background:

• Adaptor complex 3 (AP-3) is essential for intracellular trafficking from the Golgi and endosomes to late endosomal compartments.
• AP-3 dysfunction in mammals is linked to Hermansky-Pudlak syndrome type 2, cyclic neutropenias, and epileptic encephalopathy.
• AP-3 mediates cargo transport from the trans-Golgi to the lysosomal vacuole in budding yeast.

Purpose of the Study:

• To develop a rapid and efficient method for screening AP-3 mutants.
• To enable the assessment of AP-3 functional deficiency.
• To facilitate high-throughput screening for AP-3 related research.

Main Methods:

• Development of GNSI, a synthetic, genetically encoded reporter system.
• Utilizing chromogenic or growth phenotype readouts for AP-3 assessment.
• Adaptation of the system for plate array and liquid batch culture formats.

Main Results:

• The GNSI system allows for rapid plate-based assessment of AP-3 functional deficiency.
• It effectively identifies defects in both the formation and consumption of AP-3 carrier vesicles.
• The reporter system is adaptable for high-throughput screening and selection.

Conclusions:

• GNSI provides a valuable tool for studying AP-3 mediated trafficking.
• This reporter system overcomes the lack of rapid screens for AP-3 mutants.
• The availability of GNSI plasmids through Addgene repository will aid further research.