Conformational dynamics promotes disordered regions from function-dispensable to essential in evolved site-specific...

Area of Science:

• Protein intrinsically disordered regions (IDRs)
• Molecular dynamics and protein engineering

Background:

• Intrinsically disordered regions (IDRs) are crucial for molecular recognition and regulation via disorder-to-order transitions.
• Understanding structural disorder-function relationships is vital for deciphering IDR mechanisms.
• The Cre/loxP recombinase system serves as a model for studying IDR functional mechanisms.

Purpose of the Study:

• To investigate how evolution and dynamics of an intrinsically disordered N-terminal tail influence functional properties.
• To decipher the structural disorder-function relationship in the Tre/loxLTR recombinase system.
• To explore the link between protein stability and function in DNA recombination.

Main Methods:

• Utilized in vitro and in silico evolution data.
• Performed conformational dynamics studies.
• Employed AI-based folding simulations, thermodynamic stability calculations, mutagenesis, and DNA recombination assays.

Main Results:

• Discovered that partial conformational order in the Tre recombinase N-terminal tail contributes to thermodynamic stability.
• Showed that this ordered tail region packs onto a conserved hydrophobic surface.
• Established a link between protein stability and functional activity in DNA recombination.

Conclusions:

• Proposed atom-detailed mechanistic insights into disorder-function relationships.
• Highlighted the potential of N-terminal tails for regulating Cre-like tyrosine-type site-specific recombinases (SSRs).
• Suggested implications for rational engineering in biotechnology and genomic medicine.