Work

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

Year: 2004

Type: article

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.

Source: RNA

DOI: 10.1261/rna.7151404

Authors: STAVROULA MILI, JOAN A. STEITZ

Institutions: Howard Hughes Medical Institute, Yale University

Language: English

Topic: RNA Research and Splicing

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funders: Howard Hughes Medical Institute, National Institutes of Health, National Institute of General Medical Sciences

Awards: G1855224227, Vertebrate Non-Coding RNPs: Integrating function, biogenesis and survival