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Human Lin28 Forms a High-Affinity 1:1 Complex with the 106 similar to 363 Cluster miRNA miR-363

Lookup NU author(s): Dr Daniel PetersORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

Lin28A is a. post-transcriptional regulator of gene expression that interacts with and negatively regulates the biogenesis of let-7 family miRNAs. Recent data suggested that Lin28A also binds the putative tumor suppressor miR-363, a member of the 106 similar to 363 cluster of miRNAs. Affinity for this miRNA and the stoichiometry of the protein-RNA complex are unknown. Characterization of human Lin28's interaction with RNA has been complicated by difficulties in producing stable RNA-free protein. We have engineered a maltose binding protein fusion with Lin28, which binds let-7 miRNA with a K-d of 54.1 +/- 4.2 nM, in agreement with previous. data on a murine homologue. We show that human Lin28A binds miR-363 with a 1:1 stoichiometry and with a similar, if riot higher, affinity (K-d = 16.6 +/- 1.9 nM). Further analysis suggests that the interaction of the N-terminal cold shock domain of Lin28A with RNA is salt-dependent, supporting a model in which the cold shock domain allows the protein to sample RNA substrates through transient electrostatic interactions.


Publication metadata

Author(s): Peters DT, Fung HKH, Levdikov VM, Irmscher T, Warrander FC, Greive SJ, Kovalevskiy O, Isaacs HV, Coles M, Antson AA

Publication type: Article

Publication status: Published

Journal: Biochemistry

Year: 2016

Volume: 55

Issue: 36

Pages: 5021-5027

Print publication date: 13/09/2016

Online publication date: 25/08/2016

Acceptance date: 02/04/2016

Date deposited: 20/03/2017

ISSN (print): 0006-2960

ISSN (electronic): 1520-4995

Publisher: American Chemical Society

URL: http://dx.doi.org/10.1021/acs.biochem.6b00682

DOI: 10.1021/acs.biochem.6b00682


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Funding

Funder referenceFunder name
Yorkshire Cancer Research
WT095024MAWellcome Trust
WT098230Wellcome Trust

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