Mutation of the 5′-Untranslated Region Stem-Loop Structure Inhibits α1(I) Collagen Expression in Vivo

Parsons, Christopher J.; Stefanovic, Branko; Seki, Ekihiro; Aoyama, Tomonori; Latour, Anne M.; Marzluff, William F.; Rippe, Richard A.; Brenner, David A.

doi:10.1074/jbc.m110.189118

Cited by 32 publications

(33 citation statements)

References 59 publications

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“…To this end, we utilized MEFs derived from knock-in mice in which the 5ЈSL of collagen ␣1(I) mRNA was mutated (denoted here 5ЈSL Ϫ/Ϫ MEFs) (36). Immunoprecipitation with antivimentin antibody using extracts from wild-type (5ЈSL ϩ/ϩ ) MEFs pulled down collagen ␣1(I) and ␣2(I) mRNAs (Fig.…”

Section: Resultsmentioning

confidence: 99%

A Novel Role of Vimentin Filaments: Binding and Stabilization of Collagen mRNAs

Challa

Stefanovic

2011

Molecular and Cellular Biology

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The stem-loop in the 5 untranslated region (UTR) of collagen ␣1(I) and ␣2(I) mRNAs (5SL) is the key element regulating their stability and translation. Stabilization of collagen mRNAs is the predominant mechanism for high collagen expression in fibrosis. LARP6 binds the 5SL of ␣1(I) and ␣2(I) mRNAs with high affinity. Here, we report that vimentin filaments associate with collagen mRNAs in a 5SL-and LARP6-dependent manner and stabilize collagen mRNAs. LARP6 interacts with vimentin filaments through its La domain and colocalizes with the filaments in vivo. Knockdown of LARP6 by small interfering RNA (siRNA) or mutation of the 5SL abrogates the interaction of collagen mRNAs with vimentin filaments. Vimentin knockout fibroblasts produce reduced amounts of type I collagen due to decreased stability of collagen ␣1(I) and ␣2(I) mRNAs. Disruption of vimentin filaments using a drug or by expression of dominant-negative desmin reduces type I collagen expression, primarily due to decreased stability of collagen mRNAs. RNA fluorescence in situ hybridization (FISH) experiments show that collagen ␣1(I) and ␣2(I) mRNAs are associated with vimentin filaments in vivo. Thus, vimentin filaments may play a role in the development of tissue fibrosis by stabilizing collagen mRNAs. This finding will serve as a rationale for targeting vimentin in the development of novel antifibrotic therapies.Fibroproliferative disorders are leading causes of morbidity and mortality globally (1,5,11,20,28,38) and pose an enormous threat to human health. There are no effective therapies for fibrotic diseases. Excessive production of type I collagen by activated fibroblasts and myofibroblasts is the hallmark of fibroproliferative disorders. Type I collagen is a heterotrimer composed of two ␣1(I) chains and one ␣2(I) chain (29). The increased collagen synthesis can be due to the increased rate of transcription of collagen genes, increased half-life of collagen mRNAs, and their enhanced translation (33,35,47,51). Increased collagen production by activated fibroblasts is primarily due to an increase in stability of collagen mRNAs (16). During activation of hepatic stellate cells, which synthesize collagen in liver fibrosis, a 16-fold prolongation of the half-life of collagen ␣1(I) mRNAs is primarily responsible for its 50-fold-increased expression (45). The transformation of fibroblasts into myofibroblasts is also associated with increased stability of collagen ␣1(I) mRNA (37). Transforming growth factor beta (TGF-␤), the most potent profibrotic cytokine, induces collagen synthesis by prolonging the half-life of collagen ␣1(I) mRNA (51). Thus, it is now well established that stability of collagen mRNAs is the predominant mechanism regulating collagen expression (30,40,41).Two cis-acting elements were implicated in regulating the stability of collagen mRNAs. In the 3Ј untranslated region (3Ј UTR) of collagen ␣1(I) mRNA, there is a cytosine-rich sequence that interacts with the ␣CP protein; this interaction stabilizes the mRNA (32, 34). In the 5Ј UTR of col...

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Section: Resultsmentioning

confidence: 99%

A Novel Role of Vimentin Filaments: Binding and Stabilization of Collagen mRNAs

Challa

Stefanovic

2011

Molecular and Cellular Biology

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145

130

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“…Experiments were performed at cell passages 4 to 9 under serum-free conditions using a 1:1 mixture of Dulbecco's modified essential medium (DMEM) and F-12 nutrient solution (Invitrogen). Mouse embryonic fibroblasts (MEFs) from 5Ј stem-loop mutant (5ЈSL Ϫ/Ϫ ) or wild type (WT) C57/BL6 mice were obtained as previously described (7). The 5ЈSL Ϫ/Ϫ MEFs contained a 21-nt mutation in the 5Ј-UTR of the COL1a1 gene, which disrupted formation of the stem-loop structure, inhibited LARP6 binding, but did not affect the coding region of the COL1a1 gene (7,10).…”

Section: Methodsmentioning

confidence: 99%

“…Mouse embryonic fibroblasts (MEFs) from 5Ј stem-loop mutant (5ЈSL Ϫ/Ϫ ) or wild type (WT) C57/BL6 mice were obtained as previously described (7). The 5ЈSL Ϫ/Ϫ MEFs contained a 21-nt mutation in the 5Ј-UTR of the COL1a1 gene, which disrupted formation of the stem-loop structure, inhibited LARP6 binding, but did not affect the coding region of the COL1a1 gene (7,10). MEFs were cultured in 10% FCS, DMEM, and experiments were performed at passages 4 to 8.…”

Section: Methodsmentioning

confidence: 99%

“…This 5Ј stem-loop (5ЈSL) motif has been shown to regulate collagen type I mRNA expression and translation (4 -6). Mutation of the COL1a1 5ЈSL, which abolishes the stem-loop structure but does not affect the coding region of COL1a1, resulted in reduced collagen type I expression in vivo (7). In addition to promoting translation, the 5ЈSL has also been shown to be inhibitory of translation in vitro and in quiescent hepatic stellate cells in which there is an absence of protein binding to the 5ЈSL (4,6).…”

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Insulin-like Growth Factor-1 Increases Synthesis of Collagen Type I via Induction of the mRNA-binding Protein LARP6 Expression and Binding to the 5′ Stem-loop of COL1a1 and COL1a2 mRNA

Blackstock

Higashi

Sukhanov

et al. 2014

Journal of Biological Chemistry

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Background:The la ribonucleoprotein domain family member 6, LARP6, regulates collagen type 1 mRNA translation. Results: IGF-1 increases LARP6 expression, resulting in increased LARP6-collagen type 1 mRNA complex and collagen synthesis in smooth muscle. Conclusion: IGF-1 enhances collagen fibrillogenesis via induction of LARP6. Significance: This report uncovers a critical mechanism whereby IGF-1 induces a more stable plaque phenotype in atherosclerosis.

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“…Human LARP6 regulates the synthesis of collagen a1(I), a2(I), and likely a1(III) chains via a specific interaction with a unique stem-loop structure in the 59 UTR of these mRNAs that includes the initiation codon and is conserved throughout vertebrates (Cai et al 2010a,b). Recent work has shown that HsLARP6 coordinates the translation of the a1(I) and a2(I) collagen chains as well as their correct assembly, by colocalizing collagen mRNAs with intracellular nonmuscle myosin filaments whose association with polysomes controls the synthesis of heterotrimeric type I collagen (Cai et al 2010a,b;Parsons et al 2011).…”

Section: Introductionmentioning

confidence: 99%

The association of a La module with the PABP-interacting motif PAM2 is a recurrent evolutionary process that led to the neofunctionalization of La-related proteins

Merret¹,

Martino²,

Bousquet‐Antonelli³

et al. 2012

RNA

114

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La-related proteins (LARPs) are largely uncharacterized factors, well conserved throughout evolution. Recent reports on the function of human LARP4 and LARP6 suggest that these proteins fulfill key functions in mRNA metabolism and/or translation. We report here a detailed evolutionary history of the LARP4 and 6 families in eukaryotes. Genes coding for LARP4 and 6 were duplicated in the common ancestor of the vertebrate lineage, but one LARP6 gene was subsequently lost in the common ancestor of the eutherian lineage. The LARP6 gene was also independently duplicated several times in the vascular plant lineage. We observed that vertebrate LARP4 and plant LARP6 duplication events were correlated with the acquisition of a PABPinteracting motif 2 (PAM2) and with a significant reorganization of their RNA-binding modules. Using isothermal titration calorimetry (ITC) and immunoprecipitation methods, we show that the two plant PAM2-containing LARP6s (LARP6b and c) can, indeed, interact with the major plant poly(A)-binding protein (PAB2), while the third plant LARP6 (LARP6a) is unable to do so. We also analyzed the RNA-binding properties and the subcellular localizations of the two types of plant LARP6 proteins and found that they display nonredundant characteristics. As a whole, our results support a model in which the acquisition by LARP4 and LARP6 of a PAM2 allowed their targeting to mRNA 39 UTRs and led to their neofunctionalization.

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Mutation of the 5′-Untranslated Region Stem-Loop Structure Inhibits α1(I) Collagen Expression in Vivo

Cited by 32 publications

References 59 publications

A Novel Role of Vimentin Filaments: Binding and Stabilization of Collagen mRNAs

A Novel Role of Vimentin Filaments: Binding and Stabilization of Collagen mRNAs

Insulin-like Growth Factor-1 Increases Synthesis of Collagen Type I via Induction of the mRNA-binding Protein LARP6 Expression and Binding to the 5′ Stem-loop of COL1a1 and COL1a2 mRNA

The association of a La module with the PABP-interacting motif PAM2 is a recurrent evolutionary process that led to the neofunctionalization of La-related proteins

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