Synthesis and degradation of type I procollagen mRNAs in cultured human skin fibroblasts and the effect of cortisol.

Hämäläinen, Leena; Oikarinen, Jouko; Kivirikko, Kari I.

doi:10.1016/s0021-9258(20)71156-7

Cited by 144 publications

(6 citation statements)

References 32 publications

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“…Both transcriptional and posttranscriptional mechanisms may be involved in the glucocorticoid-mediated decrease of type I procollagen mRNAs. In human skin fibroblasts cortisol has been shown to increase the degradation of type I procollagen mRNAs without decreasing the synthesis of these mRNAs by nuclei in vitro (Hamalainen et al, 1985). Similar results were also reported for the effect of dexamethasone on type I procollagen mRNAs in rat dermal fibroblasts (Raghow et al, 1986).…”

Section: Discussionsupporting

confidence: 74%

Glucocorticoid coordinate regulation of type I procollagen gene expression and procollagen DNA-binding proteins in chick skin fibroblasts

Cockayne¹,

Cutroneo²

1988

Biochemistry

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Nuclei were isolated from control and dexamethasone-treated (2 h) embryonic chick skin fibroblasts and transcribed in vitro. Nuclei isolated from dexamethasone-treated fibroblasts transcribed less pro alpha 1(I) and pro alpha 2(I) mRNAs but not beta-actin mRNA. Fibroblasts receiving dexamethasone and [5,6-3H]uridine also demonstrated decreased synthesis of nuclear type I procollagen mRNAs but not beta-actin mRNA. In fibroblasts treated with cycloheximide the newly synthesized nuclear type I procollagen mRNA species were markedly decreased. An enhanced inhibitory effect was observed when fibroblasts were treated with cycloheximide plus dexamethasone. Since the studies above demonstrate that active protein synthesis is required to maintain the constitutive expression of the type I procollagen genes, we determined if glucocorticoids regulate DNA-binding proteins with sequence specificity for the alpha 2(I) procollagen gene. Nuclear protein blots were probed with the 32P-end-labeled pBR322 vector DNA and 32P-end-labeled alpha 2(I) procollagen promoter containing DNA. Nonhistone proteins remained bound to labeled DNA at stringency washes of 0.05 and 0.1 M NaCl. As the ionic strength was increased to 0.2 and 0.3 M NaCl, the nonhistone-protein DNA binding was preferentially lost. Only the low molecular weight proteins remained bound to labeled DNA at the highest ionic strength, indicating nonspecific binding of these nuclear proteins. Dexamethasone treatment resulted in an increase of binding of nonhistone proteins to vector- and promoter-labeled DNAs over that observed in control fibroblasts at stringency washes of 0.05 and 0.1 M NaCl and to a lesser extent at 0.2 M NaCl. The binding specificities of nonhistone proteins for the alpha 2(I) procollagen promoter containing DNA were calculated. Three nonhistone DNA-binding proteins of Mr 90,000, 50,000, and 30,000 had altered specificities following dexamethasone treatment.

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Section: Discussionsupporting

confidence: 74%

Glucocorticoid coordinate regulation of type I procollagen gene expression and procollagen DNA-binding proteins in chick skin fibroblasts

Cockayne¹,

Cutroneo²

1988

Biochemistry

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“…DISCUSSION Suppression of type I collagen gene expression is important in the prevention of tissue fibrosis. Previous studies have shown that glucocorticoids, as well as the cytokines IFN-y and TNF-a, are potent inhibitors of type I collagen expression, and can abrogate the enhancement of type I collagen gene expression by TGF-, [5,[7][8][9]42]. The results of the present study showed that OA, a specific inhibitor of cellular PPI and PP2A, is a potent inhibitor of expression of type I and III procollagen genes by human dermal fibroblasts.…”

Section: Suppression Of Prox2(l) Collagen Promoter Activity By Oasupporting

confidence: 63%

“…Interestingly, treatment of cells with OA in combination with TGF-,f2 prevented up-regulation of both type I and III collagen mRNA levels by TGF-,b2, resulting in reduction in these mRNA levels below the levels noted in the untreated control cells (Figure 2). Effect of OA on type I collagen mRNA levels in combination with dexamethasone Glucocorticoids potently suppress type I collagen gene expression at the post-transcriptional level [9]. We also wanted to test the effect of dexamethasone on type I collagen mRNA levels in combination with OA.…”

Section: Resultsmentioning

confidence: 99%

“…Type I collagen gene expression is potently inhibited at promoter level by tumor necrosis factor-a (TNF-a), which also abrogates the transcriptional activation of type I collagen genes by TGF-,3 [5,7]. On the other hand, interferon-y (IFN-y) and glucocorticoids potently suppress expression of type I collagen genes at post-transcriptional level [5,8,9].…”

Section: Introductionmentioning

confidence: 99%

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The protein phosphatase inhibitor okadaic acid suppresses type I collagen gene expression in cultured fibroblasts at the transcriptional level

Westermarck

Ilvonen

Kähäri

1995

Biochemical Journal

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Type I collagen is the most abundant component of the extracellular matrix of human connective tissues. We have examined the effect of okadaic acid (OA), an inhibitor of phosphoserine- and-phosphothreonine-specific protein phosphatases 1 and 2A, on type I collagen gene expression by fibroblasts in culture. Treatment of human skin fibroblasts with OA potently reduced type I and type III collagen mRNA levels, maximally by over 90%. The inhibitory effect of OA on type I and III collagen mRNA abundance was not prevented by cycloheximide, and was not affected by simultaneous treatment with dexamethasone or retinoic acid. OA also abrogated the enhancing effect of transforming growth factor-beta (TGF-beta) on type I and III collagen mRNA levels. Treatment of transiently transfected NIH-3T3 fibroblasts with OA suppressed the activity of a 3.5 kb human pro alpha 2(I) collagen promoter/chloramphenicol acetyltransferase construct maximally, by 70%. In addition, OA treatment of NIH-3T3 cells abrogated enhancement of pro alpha 2(I) collagen promoter activity by TGF-beta. These results indicate that protein phosphatases 1 and 2A have an important role as positive regulators of type I and III collagen gene expression. The results also suggest that selective inhibition of activity of protein phosphatases 1 and 2A may offer a novel approach for preventing excessive collagen accumulation in fibrotic disorders.

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“…For these reasons, the values for expression of a type I collagen gene based on reporter gene constructs are difficult to relate in a quantitative manner to expression of the normal gene in transgenic animals. Similar problems are encountered in cell transfection experiments because the expression of type I procollagen genes in cultured fibroblasts varies widely with the state of confluency of the cells, the passage number, and other conditions of cell culture [see Hamalainen et al (1985) and Olsen and Prockop (1989)]. As reported previously (Olsen et al, 1991), the human minigene constructs made it possible to assay expression of the exogenous gene relative to the endogenous genes in stably transfected 3T3 in cells.…”

Section: Discussionmentioning

confidence: 95%

Tissue- and development-specific expression in transgenic mice of a type I procollagen (COL1A1) minigene construct with 2.3 kb of the promoter region and 2 kb of the 3'-flanking region. Specificity is independent of the putative regulatory sequences in the first intron

Sokolov¹,

Mays²,

Khillan³

et al. 1993

Biochemistry

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Previous reports have provided inconsistent data as to the cis-regulatory elements that are essential for correct expression of the gene for the pro alpha 1 (I) chain of type I procollagen (COL1A1) in the many tissues in which the protein is synthesized. Here, two internally deleted minigene versions of the human COL1A1 gene were used to prepare transgenic mice. The constructs made it possible to test regulatory sequences in the normal context of the gene. Also, in contrast to the reporter genes used in previous experiments, the constructs made it possible to assay quantitatively expression of the exogenous genes relative to expression of the endogenous COL1A1 gene, both as mRNA and as protein. The average level of expression of the minigenes varied among three transgenic lines, but the ratio of expression of the minigenes to expression of the endogenous gene was the same in all transgenic mice of a given line. Within the same line, the ratio of expression was essentially the same in nine or more tissues in which expression of the endogenous gene varied widely. Also, the ratio of expression within a given line was the same in 15-day-old embryos and in mice ranging in age from 4 days to 4 months. In addition, the ratio remained constant during repair of a surgical wound. The results demonstrated, therefore, that the minigene constructs with about 2.3 kb of the promoter region and about 2 kb of the 3'-flanking region contained all of the sequences necessary for correct expression of the genes in a tissue-specific and development-specific manner.(ABSTRACT TRUNCATED AT 250 WORDS)

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Synthesis and degradation of type I procollagen mRNAs in cultured human skin fibroblasts and the effect of cortisol.

Cited by 144 publications

References 32 publications

Glucocorticoid coordinate regulation of type I procollagen gene expression and procollagen DNA-binding proteins in chick skin fibroblasts

Glucocorticoid coordinate regulation of type I procollagen gene expression and procollagen DNA-binding proteins in chick skin fibroblasts

The protein phosphatase inhibitor okadaic acid suppresses type I collagen gene expression in cultured fibroblasts at the transcriptional level

Tissue- and development-specific expression in transgenic mice of a type I procollagen (COL1A1) minigene construct with 2.3 kb of the promoter region and 2 kb of the 3'-flanking region. Specificity is independent of the putative regulatory sequences in the first intron

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