A Conserved Stem Loop Motif in the 5′Untranslated Region Regulates Transforming Growth Factor-β1 Translation
Open Access
- 26 August 2010
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 5 (8), e12283
- https://doi.org/10.1371/journal.pone.0012283
Abstract
Transforming growth factor-β1 (TGF-β1) regulates cellular proliferation, differentiation, migration, and survival. The human TGF-β1 transcript is inherently poorly translated, and translational activation has been documented in relation to several stimuli. In this paper, we have sought to identify in cis regulatory elements within the TGF-β1 5′Untranslated Region (5′UTR). In silico analysis predicted formation of stable secondary structure in a G/C-rich element between nucleotides +77 to +106, and demonstrated that this element is highly conserved across species. Circular dichroism spectroscopy confirmed the presence of secondary structure in this region. The proximal 5′UTR was inhibitory to translation in reporter gene experiments, and mutation of the secondary structure motif increased translational efficiency. Translational regulation of TGF-β1 mRNA is linked to altered binding of YB-1 protein to its 5′UTR. Immunoprecipitation-RT-qPCR demonstrated a high basal association of YB-1 with TGF-β1 mRNA. However, mutation of the secondary structure motif did not prevent interaction of YB-1 with the 5′UTR, suggesting that YB-1 binds to this region due to its G/C-rich composition, rather than a specific, sequence-dependent, binding site. These data identify a highly conserved element within the TGF-β1 5′UTR that forms stable secondary structure, and is responsible for the inherent low translation efficiency of this cytokine.This publication has 59 references indexed in Scilit:
- General RNA-binding proteins have a function in poly(A)-binding protein-dependent translationThe EMBO Journal, 2008
- The DEAD-Box RNA Helicase DDX3 Associates with Export Messenger Ribonucleoproteins as well asTip-associated Protein and Participates in Translational ControlMolecular Biology of the Cell, 2008
- Role of transforming growth factor-β superfamily signaling pathways in human diseaseBiochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2008
- Y-box protein-1 controls transforming growth factor-β1 translation in proximal tubular cellsKidney International, 2008
- The mRNA-binding Protein YB-1 (p50) Prevents Association of the Eukaryotic Initiation Factor eIF4G with mRNA and Inhibits Protein Synthesis at the Initiation StagePublished by Elsevier BV ,2003
- Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT MethodMethods, 2001
- The Major Messenger Ribonucleoprotein Particle Protein p50 (YB-1) Promotes Nucleic Acid Strand AnnealingPublished by Elsevier BV ,2001
- HK-2: An immortalized proximal tubule epithelial cell line from normal adult human kidneyKidney International, 1994
- The equilibrium partition function and base pair binding probabilities for RNA secondary structurePeptide Science, 1990
- Production of transforming growth factor beta by human T lymphocytes and its potential role in the regulation of T cell growth.The Journal of Experimental Medicine, 1986