Evidence for reassociation of RNA-binding proteins after cell lysis: Implications for the interpretation of immunoprecipitation analyses

2004/09/23 by Stavroula Mili, Joan A. Steitz · 13 citations
Biochemistry, Genetics and Molecular Biology · #RNA Research and Splicing #RNA and protein synthesis mechanisms #RNA modifications and cancer #Immunoprecipitation #Ribonucleoprotein #Biology #RNA #Lysis #RNA-binding protein #Cell biology #Chromatin immunoprecipitation #Molecular biology #Computational biology #Biochemistry #Gene #Gene expression

paper · pdf · doi:10.1261/rna.7151404

openalex publication_date 2004/09/23 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/01

Abstract

Immuno- and other affinity-purification approaches are commonly used to characterize the composition of ribonucleoprotein complexes. While associations detected by these procedures are often interpreted as reflecting in vivo interactions, it is also possible that they arise from reassociation of molecules after cell lysis. Here we used an experimental approach that allowed us to distinguish between these possibilities. Surprisingly, we show that the association of the RNA-binding protein HuR with its target mRNA, c-fos, as detected by co-immunoprecipitation, results largely from reassociation of molecules subsequent to cell lysis. The existence of such postlysis reassortments thus demonstrates that co-immunoprecipitation does not always recapitulate the in vivo state of ribonucleoprotein complexes.

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