Teashirt and C-Terminal Binding Protein Interact to Regulate Drosophila Eye Development

Area of Science:

• Developmental Biology
• Genetics
• Molecular Biology

Background:

• The Drosophila retinal determination network involves Teashirt (Tsh) and C-terminal Binding Protein (CtBP), crucial for eye development.
• Tsh and CtBP are co-expressed in proliferating eye precursor cells within the larval eye-antennal disc.
• Previous studies suggested genetic interaction and molecular complex formation, but in vivo interaction in eye discs was unknown.

Purpose of the Study:

• To investigate whether Teashirt (Tsh) and C-terminal Binding Protein (CtBP) interact within Drosophila eye-antennal discs.
• To determine if this interaction affects adult eye development.
• To elucidate the molecular and genetic mechanisms underlying this interaction.

Main Methods:

• Generated transgenic Drosophila strains with 5' GFP-tagged tsh and 5' FLAG-tagged ctbp using CRISPR-Cas9 and BAC recombineering.
• Verified protein expression via immunohistology and confocal microscopy in larval eye-antennal discs.
• Employed genetic screening, Scanning Electron Microscopy, co-immunoprecipitation, and GST pulldown assays to assess interaction and effects.

Main Results:

• Demonstrated that Teashirt (Tsh) and C-terminal Binding Protein (CtBP) interact both genetically and physically in the third instar larval eye-antennal disc.
• This interaction plays a role in regulating adult eye development.
• The interaction appears to limit the proliferation of eye precursor cells in the larval eye disc.

Conclusions:

• Teashirt (Tsh) and C-terminal Binding Protein (CtBP) physically and genetically interact within the Drosophila third instar larval eye-antennal disc.
• The relative abundance of Tsh and CtBP influences their interaction's outcome on Drosophila eye formation.
• This interaction is critical for controlling eye precursor cell populations and ensuring proper adult eye development.