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Single-molecule approach to immunoprecipitated protein complexes: Insights into miRNA uridylation

Cited 58 time in Web of Science Cited 61 time in Scopus
Issue Date
2011-07
Citation
EMBO Reports, Vol.12 No.7, pp.690-696
Keywords
single-molecule fluorescencesingle-molecule immunoprecipitationTUT4 (ZCCHC11)Lin28 (Lin28a, Lin28b, Lin28)real-time hybridization
Abstract
Single-molecule techniques have been used for only a subset of biological problems because of difficulties in studying proteins that require cofactors or post-translational modifications. Here, we present a new method integrating single-molecule fluorescence microscopy and immunopurification to study protein complexes. We used this method to investigate Lin28-mediated microRNA uridylation by TUT4 (terminal uridylyl transferase 4, polyU polymerase), which regulates let-7 microRNA biogenesis. Our real-time analysis of the uridylation by the TUT4 immunoprecipitates suggests that Lin28 functions as a processivity factor of TUT4. Our new technique, SIMPlex (single-molecule approach to immunoprecipitated protein complexes), provides a universal tool to analyse complex proteins at the single-molecule level.
ISSN
1469-221X
URI
https://hdl.handle.net/10371/171890
DOI
https://doi.org/10.1038/embor.2011.100
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College of Natural Sciences (자연과학대학)Dept. of Biological Sciences (생명과학부)Journal Papers (저널논문_생명과학부)
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