S6K2-mediated regulation of TRBP as a determinant of miRNA expression in human primary lymphatic endothelial cells

TY - JOUR

T1 - S6K2-mediated regulation of TRBP as a determinant of miRNA expression in human primary lymphatic endothelial cells

AU - Warner, Matthew J

AU - Bridge, Katherine S

AU - Hewitson, James P

AU - Hodgkinson, Michael R

AU - Heyam, Alex

AU - Massa, Bailey C

AU - Haslam, Jessica C

AU - Chatzifrangkeskou, Maria

AU - Evans, Gareth J O

AU - Plevin, Michael J

AU - Sharp, Tyson V

AU - Lagos, Dimitris

N1 - Funding Information: We would like to thank Banushan Balansethupathy for technical contributions. Wellcome Trust [097829; through the Centre for Chronic Diseases and Disorders at the University of York]; Medical Research Council [MR/L008505/1]; Yorkshire Cancer Research [TPP053 to D.L.]; BBSRC [BB/I007571/2 to T.V.S.]. 'Combating Infectious Disease: Computational Approaches in Translational Science' Wellcome Trust PhD programme [WT095024MA to A.P.H.]. Funding for open access charge: The University of York Open Access Fund through agreement with the Medical Research Council and The Wellcome Trust who funded the work. Conflict of interest statement. None declared. Publisher Copyright: © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. Copyright: Copyright 2017 Elsevier B.V., All rights reserved.

PY - 2016/11/16

Y1 - 2016/11/16

N2 - MicroRNAs (miRNAs) are short non-coding RNAs that silence mRNAs. They are generated following transcription and cleavage by the DROSHA/DGCR8 and DICER/TRBP/PACT complexes. Although it is known that components of the miRNA biogenesis machinery can be phosphorylated, it remains poorly understood how these events become engaged during physiological cellular activation. We demonstrate that S6 kinases can phosphorylate the extended C-terminal domain of TRBP and interact with TRBP in situ in primary cells. TRBP serines 283/286 are essential for S6K-mediated TRBP phosphorylation, optimal expression of TRBP, and the S6K-TRBP interaction in human primary cells. We demonstrate the functional relevance of this interaction in primary human dermal lymphatic endothelial cells (HDLECs). Angiopoietin-1 (ANG1) can augment miRNA biogenesis in HDLECs through enhancing TRBP phosphorylation and expression in an S6K2-dependent manner. We propose that the S6K2/TRBP node controls miRNA biogenesis in HDLECs and provides a molecular link between the mTOR pathway and the miRNA biogenesis machinery.

AB - MicroRNAs (miRNAs) are short non-coding RNAs that silence mRNAs. They are generated following transcription and cleavage by the DROSHA/DGCR8 and DICER/TRBP/PACT complexes. Although it is known that components of the miRNA biogenesis machinery can be phosphorylated, it remains poorly understood how these events become engaged during physiological cellular activation. We demonstrate that S6 kinases can phosphorylate the extended C-terminal domain of TRBP and interact with TRBP in situ in primary cells. TRBP serines 283/286 are essential for S6K-mediated TRBP phosphorylation, optimal expression of TRBP, and the S6K-TRBP interaction in human primary cells. We demonstrate the functional relevance of this interaction in primary human dermal lymphatic endothelial cells (HDLECs). Angiopoietin-1 (ANG1) can augment miRNA biogenesis in HDLECs through enhancing TRBP phosphorylation and expression in an S6K2-dependent manner. We propose that the S6K2/TRBP node controls miRNA biogenesis in HDLECs and provides a molecular link between the mTOR pathway and the miRNA biogenesis machinery.

KW - Angiopoietin-1/pharmacology

KW - Cell Line

KW - Cells, Cultured

KW - Endothelial Cells/drug effects

KW - Gene Expression Regulation/drug effects

KW - Humans

KW - MicroRNAs/genetics

KW - Phosphorylation

KW - Protein Interaction Domains and Motifs

KW - RNA-Binding Proteins/chemistry

KW - Ribosomal Protein S6 Kinases/metabolism

UR - http://www.scopus.com/inward/record.url?scp=85016107462&partnerID=8YFLogxK

U2 - 10.1093/nar/gkw631

DO - 10.1093/nar/gkw631

M3 - Article

C2 - 27407113

SN - 0305-1048

VL - 44

SP - 9942

EP - 9955

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 20

ER -