Related Experiment Video
Updated: May 9, 2026
08:14
In Vitro Biochemical Assays using Biotin Labels to Study Protein-Nucleic Acid Interactions
Published on: July 17, 2019
Experimental characterization of the human non-sequence-specific nucleic acid interactome
Genome Biology
|August 2, 2013
Summary
This study systematically identified human proteins that bind nucleic acids, discovering 746 direct binders, including novel ones. It reveals protein-nucleic acid interactions and YB-1
Area of Science:
- Molecular Biology
- Proteomics
- Genomics
Background:
- Protein-nucleic acid interactions are crucial for gene transcription, RNA homeostasis, protein translation, and innate immunity.
- Previous studies focused on mRNA-binding proteins, leaving a gap in understanding non-sequence-specific interactions with various nucleic acids.
Purpose of the Study:
- To systematically investigate the non-sequence-specific binding of native human proteins to diverse nucleic acids, including methylated cytosine.
- To identify and profile proteins interacting with RNA and DNA using a comprehensive experimental approach.
Main Methods:
- Developed an affinity purification mass spectrometry and bioinformatics workflow.
- Utilized 25 rationally designed nucleic acid probes against whole cell extracts from three human cell lines.
- Profiled protein binding preferences across different nucleic acid types.
Main Results:
- Identified 746 high-confidence direct nucleic acid-binding proteins.
- Discovered 139 novel nucleic acid-binding proteins and 237 with no prior experimental evidence.
- Assigned specific binding affinities to sub-types of nucleic acid probes for 219 proteins and domains, including YB-1's preferential binding to methylated cytosine.
Conclusions:
- The study provides a valuable dataset of experimentally determined nucleic acid-binding proteins.
- The methodology offers significant potential for future research at the protein-nucleic acid interface.
