ECT2 peptide sequences outside the YTH domain regulate its m⁶A-RNA binding.

Area of Science:

• Molecular Biology
• Genetics
• Biochemistry

Background:

• N6-methyladenosine (m6A) is a prevalent RNA modification crucial for eukaryotic gene expression.
• YTH domain proteins are key readers of m6A modifications, but their regulatory mechanisms are not fully understood.
• Intrinsically Disordered Regions (IDRs) in YTH proteins, like Arabidopsis thaliana's ECT2, are implicated in functional specialization.

Purpose of the Study:

• To biochemically characterize the m6A binding properties of the ECT2 protein.
• To identify regulatory regions outside the YTH domain that influence m6A binding.
• To investigate factors enhancing ECT2's selectivity for m6A-modified RNA.

Main Methods:

• In vitro biochemical characterization of full-length ECT2 and its YTH domain.
• Analysis of m6A-methylated RNA binding affinity and specificity.
• Identification of regulatory peptide regions and structural elements.

Main Results:

• Full-length ECT2 and its YTH domain exhibit distinct m6A binding capabilities.
• N-terminal IDR regions of ECT2 regulate its binding to m6A-methylated RNA.
• ECT2's m6A binding selectivity is enhanced by flanking uridine-rich sequences.
• Conserved structural elements near the YTH domain further enhance m6A binding.

Conclusions:

• Novel regulatory regions outside the YTH domain control ECT2's m6A RNA binding.
• Uridine content and specific structural elements modulate ECT2's binding affinity and selectivity.
• These regulatory mechanisms may be conserved across eukaryotic YTH readers.