Integrative structural biology of protein-RNA interactions in four dimensions

Area of Science:

• Structural biology
• Molecular biology
• Biochemistry

Background:

• Protein-RNA complexes are crucial for cellular functions.
• Their inherent flexibility, multi-valency, and dynamics pose significant challenges for traditional structural analysis.
• Understanding the structure and dynamics of ribonucleoprotein (RNP) assemblies is essential for deciphering cellular mechanisms.

Purpose of the Study:

• To review recent advancements in integrative modeling of protein-RNA complexes.
• To highlight progress in in-cell, 4D, and condensate structural biology.
• To discuss how these integrative approaches enhance our understanding of RNP assembly, regulation, and function in cellular environments.

Main Methods:

• Integrative structural biology approaches combining NMR, small-angle scattering, cryo-EM, cryo-ET, crosslinking mass spectrometry, and single-molecule techniques.
• Artificial intelligence (AI)-based predictions for modeling complex RNP assemblies.
• Characterization of RNP assemblies both in vitro and within their native cellular contexts.

Main Results:

• Successful characterization of increasingly complex RNP assemblies using integrative strategies.
• Enabled capture of molecular architecture and dynamic behaviors across time and space, leading to 4D structural biology.
• Significant advances in studying RNPs in cellular environments and as condensates.

Conclusions:

• Integrative modeling is revolutionizing the structural analysis of challenging protein-RNA complexes.
• These multi-technique approaches provide unprecedented insights into RNP assembly, regulation, and function.
• The development of 4D structural biology and in-cell structural studies is crucial for understanding cellular processes.