RNA targets of wild-type and mutant FET family proteins

Area of Science:

• Molecular Biology
• Genetics
• Neuroscience

Background:

• The FET (FUS, EWSR1, TAF15) protein family are conserved RNA-binding proteins implicated in oncogenesis and neuronal diseases.
• Despite their known roles, the specific RNA targets and recognition elements of FET proteins remain largely unknown.

Purpose of the Study:

• To globally define the RNA targets for all human FET proteins using a high-throughput method.
• To investigate how mutations in FUS, a member of the FET family, affect its RNA binding profiles and subcellular localization.

Main Methods:

• Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) was employed to identify global RNA targets.
• Analysis of RNA binding patterns for wild-type FET proteins and two Amyotrophic Lateral Sclerosis (ALS)-associated FUS mutants.

Main Results:

• PAR-CLIP successfully identified global RNA targets for human FET proteins.
• FET family members (FUS, EWSR1, TAF15) exhibited similar RNA binding profiles.
• FUS mutants associated with ALS displayed significantly altered RNA binding patterns compared to wild-type FUS.
• The observed changes in FUS mutant binding patterns correlated with alterations in their subcellular localization.

Conclusions:

• This study elucidates the global RNA interactomes of FET proteins, providing critical insights into their functions.
• FUS mutations disrupt normal RNA binding, offering a molecular basis for their role in ALS pathogenesis.
• Understanding FET protein-RNA interactions is crucial for deciphering their roles in health and disease.