Transforming growth factor-beta reverses a posttranscriptional defect in elastin synthesis in a cutis laxa skin fibroblast strain.

Mc, Zhang; Giro, M.G.; Quaglino, Daniela; Davidson, J.M.

doi:10.1172/jci117808

Cited by 42 publications

(24 citation statements)

References 40 publications

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“…(24) and above for WM and WS, TGF-␤ could in part restore tropoelastin expression in these cutis laxa fibroblasts by stabilizing ELN mRNA. To examine whether partial restoration of ELN mRNA stability was functioning through selective expression of ELN mRNA from the normal allele, RT-PCR products amplifying the region spanning nucleotides 1671-2292 from TGF-␤-treated and untreated K.T.…”

Section: Tgf-␤ Does Not Change the Ratio Of Eln Mrna Expressed From Twomentioning

confidence: 86%

“…Histologic, ultrastructural, and biochemical analyses of skin and cultured fibroblasts from the patient have been reported previously (17,24). Briefly, dermal collagen fibers appear normal, but elastic fibers appear fragmented with a paucity of amorphous elastin in the matrix.…”

Section: Methodsmentioning

confidence: 95%

“…We have previously shown in one cutis laxa cell strain that transcript instability was the basis of a defect in ELN mRNA accumulation and tropoelastin production (24). TGF-␤ was able to increase mRNA stability markedly and to stimulate production of immunoreactive tropoelastin protein in this cell strain.…”

mentioning

confidence: 84%

“…RNA Stability-Elastin production and mRNA stability studies were carried out as described previously (17,24). RNA was isolated and purified using the QIAshredder TM and the RNeasy TM mini kit (QIA-GEN Inc., Chatsworth, CA) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

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Cutis Laxa Arising from Frameshift Mutations in Exon 30 of the Elastin Gene (ELN)

Zhang¹,

He²,

Giro³

et al. 1999

Journal of Biological Chemistry

164

114

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Congenital cutis laxa, a rare syndrome with marked skin laxity and pulmonary and cardiovascular compromise, is due to defective elastic fiber formation. In several cases, skin fibroblast tropoelastin production is markedly reduced yet reversed in vitro by transforming growth factor-␤ treatment. We previously showed that this reversal was due to elastin mRNA stabilization in one cell strain, and here this behavior was confirmed in skin fibroblasts from two generations of a second family. cDNA sequencing and heteroduplex analysis of elastin gene transcripts from three fibroblast strains in two kindreds now identify two frameshift mutations (2012⌬G and 2039⌬C) in elastin gene exon 30, thus leading to missense C termini. No other mutations were present in the ELN cDNA sequences of all three affected individuals. Transcripts from both alleles in each kindred were unstable and responsive to transforming growth factor-␤. Exons 22, 23, 26A, and 32 were always absent. Since exon 30 underwent alternative splicing in fibroblasts, we speculate that a differential splicing pattern could conceivably lead to phenotypic rescue. These two dominant-acting, apparently de novo mutations in the elastin gene appear to be responsible for qualitative and quantitative defects in elastin, resulting in the cutis laxa phenotype.

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Section: Tgf-␤ Does Not Change the Ratio Of Eln Mrna Expressed From Twomentioning

confidence: 86%

Section: Methodsmentioning

confidence: 95%

mentioning

confidence: 84%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Cutis Laxa Arising from Frameshift Mutations in Exon 30 of the Elastin Gene (ELN)

Zhang¹,

He²,

Giro³

et al. 1999

Journal of Biological Chemistry

164

114

View full text Add to dashboard Cite

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“…To assess the effect of iron on elastin mRNA stability, parallel quadruplicate cultures were grown to confluency in 100-mm dishes. Media were then changed, supplemented with 60 M transcription blocker, DRB (51), and cultures were maintained in the presence or absence of 20 and 200 M FAC for 0, 6, 12, and 24 h. Total RNA was extracted using the RNeasy Mini kit, according to manufacturer's instructions, at indicated time points. The reverse transcriptase reaction was performed using 1.5 g of total RNA, oligo(dT)s, and the SuperScript First-Strand synthesis system (Invitrogen) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Fluctuations of Intracellular Iron Modulate Elastin Production

Bunda

Kaviani

Hinek

2005

Journal of Biological Chemistry

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Production of insoluble elastin, the major component of elastic fibers, can be modulated by numerous intrinsic and exogenous factors. Because patients with hemolytic disorders characterized with fluctuations in iron concentration demonstrate defective elastic fibers, we speculated that iron might also modulate elastogenesis. In the present report we demonstrate that treatment of cultured human skin fibroblasts with low concentration of iron 2-20 M (ferric ammonium citrate) induced a significant increase in the synthesis of tropoelastin and deposition of insoluble elastin. Northern blot and realtime reverse transcription-PCR analysis revealed that treatment with 20 M iron led to an increase of ϳ3-fold in elastin mRNA levels. Because treatment with an intracellular iron chelator, desferrioxamine, caused a significant decrease in elastin mRNA level and consequent inhibition of elastin deposition, we conclude that iron facilitates elastin gene expression. Our experimental evidence also demonstrates the existence of an opposite effect, in which higher, but not cytotoxic concentrations of iron (100 -400 M) induced the production of intracellular reactive oxygen species that coincided with a significant decrease in elastin message stability and the disappearance of iron-dependent stimulatory effect on elastogenesis. This stimulatory elastogenic effect was reversed, however, in cultures simultaneously treated with high iron concentration (200 M) and the intracellular hydroxyl radical scavenger, dimethylthiourea. Thus, presented data, for the first time, demonstrate the existence of two opposite iron-dependent mechanisms that may affect the steady state of elastin message. We speculate that extreme fluctuations in intracellular iron levels result in impaired elastic fiber production as observed in hemolytic diseases.Mature elastic fibers and laminae provide extensibility and elastic recoil to vascular walls and ligaments and form a connective tissue framework of lungs, elastic cartilage, and skin (1, 2). They are complex structures made of polymeric (insoluble) elastin and 12-nm microfibrils that consist of several glycoproteins, e.g. fibrillins, fibulins, and microfibril-associated glycoproteins (3-6). Elastic fiber formation (elastogenesis) is a complex process involving several intracellular and extracellular events. Cells (fibroblasts, endothelial cells, chondroblasts, or vascular smooth muscle cells) must first synthesize and secrete numerous glycoproteins to form a microfibrillar scaffold upon which tropoelastin, the precursor peptides, are properly assembled and covalently cross-linked by lysyl oxidase into a resilient polymer, insoluble elastin (7-10). Production of elastin reaches its highest levels in the third trimester of the fetal life and steadily decreases during early postnatal development (11,12). In undisturbed tissues, elastic fibers may last over the entire human lifespan (13,14).The net deposition of elastin appears to be controlled on both the transcriptional level (tropoelastin mRNA message express...

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Transcriptional and post-transcriptional control of lysyl oxidase expression in vascular smooth muscle cells: Effects of TGF-β1 and serum deprivation

et al. 1997

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Transforming growth factor-beta 1 (TGF-beta 1) markedly reduced cell proliferation and elevated steady state lysyl oxidase (LO) mRNA 3-fold in neonatal rat aorta smooth muscle cells cultured in medium containing 10% fetal bovine serum. The increase in LO mRNA was prevented by the presence of cycloheximide, indicative of controlling events at the level of protein synthesis. The basal level of mRNA in cells proliferating in 10% fetal bovine serum in the absence of TGF-beta 1 was enhanced 7-fold upon decreasing growth by shifting to medium containing 0.5% serum. Changes in LO activity paralleled those in LO mRNA. Nuclear run-on assays revealed that the stimulation of expression in 0.5% serum involved increased gene transcription whereas that caused by TGF-beta 1 was mostly post-transcriptional in origin. LO mRNA was quite labile (t1/2 approximately 3 h) in 10% serum but was markedly stabilized (t1/2 > 12 h) by the presence of TGF-beta 1 in the 10% serum medium. LO mRNA was also considerably more stable under retarded growth conditions (0.5% serum) in the absence of TGF-beta 1. LO promoter activity in luciferase reporter constructs transfected into these cells was low and not significantly affected by the addition of TGF-beta 1 to the 10% serum medium but was markedly elevated by shifting from 10 to 0.5% serum in the absence of TGF-beta 1. Thus, LO expression is inversely correlated with cell proliferation, and is subject to control at transcriptional and post-transcriptional levels. TGF-beta 1 enhances LO expression in these cells by dramatically stabilizing LO mRNA.

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Transforming growth factor-beta reverses a posttranscriptional defect in elastin synthesis in a cutis laxa skin fibroblast strain.

Cited by 42 publications

References 40 publications

Cutis Laxa Arising from Frameshift Mutations in Exon 30 of the Elastin Gene (ELN)

Cutis Laxa Arising from Frameshift Mutations in Exon 30 of the Elastin Gene (ELN)

Fluctuations of Intracellular Iron Modulate Elastin Production

Transcriptional and post-transcriptional control of lysyl oxidase expression in vascular smooth muscle cells: Effects of TGF-β1 and serum deprivation

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