Retrovirus-induced insertional mutation in Mov13 mice affects collagen I expression in a tissue-specific manner

Kratochwil, Klaus; Mark, Klaus von der; Kollar, Edward J.; Jaenisch, Rudolf; Mooslehner, Katrin A.; Schwarz, Michaela; Haase, Kirsten; Gmachl, Ilse; Harbers, Klaus

doi:10.1016/0092-8674(89)90795-2

Cited by 110 publications

(69 citation statements)

References 40 publications

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“…Interestingly, therefore, the Movl3 phenotype does not appear to result from a generalized and uniform reduction of transcriptional activity. This heterogeneity contrasts with what we had found in the tooth where the mutant allele is expressed throughout the odontoblast layer (Kratochwil et al, 1989;Schwarz et al, 1990).…”

Section: Limited Expression Of the Movl3 Allele Leads To Impaired Boncontrasting

confidence: 97%

“…We have suggested that the differences between odontoblast and fibroblast reflect the use of different cis-acting regulatory elements in the 5' upstream region and in the first intron of the collagen gene (Kratochwil et al, 1989; see also Ramirez and DiLiberto, 1990;Pavlin et al, 1992). The position of the mutagenic insert would argue for strong upstream enhancers in the less affected cell types that allow them to overcome the transcriptional block.…”

Section: Tissue-specific Differences In the Expression Of The Movl3 Amentioning

confidence: 99%

“…Transcription of this mutant Collal allele is completely blocked in all mesodermal cell types tested so far, with the exception of the odontoblast where it is expressed at an apparently normal rate (Kratochwil et al [1989] Cell 5E807-816). To define the tissue specificity of the mutant allele more precisely, we have now studied its expression in osteoblasts, another skeletogenic cell type which, like odontoblasts, produces high amounts of collagen I.…”

Section: Introductionmentioning

confidence: 98%

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Restricted expression of Mov13 mutant α1(I) collagen gene in osteoblasts and its consequences for bone development

Kratochwil¹,

Ghaffari-Tabrizi²,

Holzinger³

et al. 1993

Developmental Dynamics

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Cell type-specific differences in the transcriptional control of the mouse gene coding for the a1 chain of collagen type I (Collal) have been revealed previously with the help of the Movl3 mouse strain which carries a retroviral insert in the first intron of the gene. Transcription of this mutant Collal allele is completely blocked in all mesodermal cell types tested so far, with the exception of the odontoblast where it is expressed at an apparently normal rate (Kratochwil et al. [1989] Cell 5E807-816). To define the tissue specificity of the mutant allele more precisely, we have now studied its expression in osteoblasts, another skeletogenic cell type which, like odontoblasts, produces high amounts of collagen I. Evidence for transcription of the Movl3 allele was obtained by in situ hybridization in homozygous (M/M) and heterozygous (M/ + ) bone tissue, in grafts as well as in vivo. The presence of mouse collagen I and the development of bone tissue were demonstrated in M/M skeletal elements grown on the chick chorioallantoic membrane (CAM). Further support for expression of the mutant gene was obtained from two 16 day MIM fetuses in vivo. Bone tissue of diverse embryological origin (vertebrae and ribs of somitic origin, long bones derived from lateral plate, calvariae from head paraxial mesoderm, and mandibulae from head neural crest) expresses the mutant allele. However, in situ hybridization experiments indicate that only a subpopulation of osteoblasts is capable of transcribing it at a high rate, resulting in severe impairment of bone development in grafts and in vivo. Therefore, osteoblasts, in comparison to odontoblasts and fibroblast-like cells, assume an intermediate position with respect to transcription of the Movl3 allele. We suggest that this diversity in the utilization of the mutant collagen gene reflects cell type-specific differences in the transcriptional regulation of the wild type (wt) Collal gene.

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Section: Limited Expression Of the Movl3 Allele Leads To Impaired Boncontrasting

confidence: 97%

Section: Tissue-specific Differences In the Expression Of The Movl3 Amentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

See 1 more Smart Citation

Restricted expression of Mov13 mutant α1(I) collagen gene in osteoblasts and its consequences for bone development

Kratochwil¹,

Ghaffari-Tabrizi²,

Holzinger³

et al. 1993

Developmental Dynamics

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“…The synthesis and deposition of ECM proteins is highly regulated and plays a critical role in tissue development, homeostasis, wound healing, and inflammation (1)(2)(3). Overexpression of ECM proteins may lead to tissue fibrosis with severe impairment of organ function.…”

Section: Introductionmentioning

confidence: 99%

Mithramycin selectively inhibits collagen-alpha 1(I) gene expression in human fibroblast.

Nehls¹,

Brenner²,

Gruss³

et al. 1993

J. Clin. Invest.

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The products of the collagen-al (I) and -a2(I) genes form the triple helical molecule collagen type I, which constitutes the major ECM protein in tissue fibrosis. The collagen-al (I) gene is mainly transcriptionally regulated, and its promoter activity depends on the interaction of the transcription factors NF-I and Spi with a tandem repeat of evolutionary conserved NF-I/Spl switch elements. An increased affinity of Spl to these elements has been observed in experimental liver fibrosis. Here, we demonstrate that the DNA binding drug mithramycin displays a high affinity binding to the GC-rich elements in the collagenal (I) promoter as measured by DNAse I protection and gel retardation assays.Mithramycin interferes with Spl but not with NF-I binding to these sites. At a concentration of 100 nM, mithramycin efficiently reduces basal and TGF--stimulated al (I) gene expression in human primary fibroblasts. The transcriptional activity and mRNA steady state levels of other genes, including the collagenase gene, as well as the growth rate of fibroblasts remained unchanged on exposure to this drug. Taken together, our results indicate that the transcriptional activity of the type I collagen gene highly depends on its GC-rich regulatory elements, and further, that these elements can be differentially blocked, thereby changing the balance between ECM structural and degrading gene activities in human fibroblasts. (J.

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“…In at least two cases provirus insertion disrupts expression of cellular genes in a tissue-specific manner (35,36). A distinct feature of the A6 mutant is the fact that provirus integration results only in a partial inactivation of the UbcM4 gene: in all tissues of homozygous embryos studied so far the level of steady-state mRNA is reduced by about 70% when analyzed on Northern blots.…”

Section: Discussionmentioning

confidence: 98%

Provirus integration into a gene encoding a ubiquitin-conjugating enzyme results in a placental defect and embryonic lethality.

Harbers

Müller

Grams

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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(2). Very recent evidence suggests that ubiquitination of membrane-bound receptors might also serve as a signal for degradation via the lysosomal pathway (4). Although ubiquitin conjugation is mainly thought to tag proteins for destruction, several lines of evidence suggest that it might also be involved in other cellular functions. This is indicated, for example, by the presence of stable ubiquitinated proteins, such as histones (5), and by the existence of enzymes that remove ubiquitin from conjugates and therefore make this modification reversible (6). Reversible ubiquitination might have a role in modulating the function of target proteiIns similar to protein phosphorylation. Attachment of ubiquitin to cellular proteins is carried out by the sequential action of three classes of enzymes, ubiquitinactivating enzymes (referred to as El or Uba), ubiquitinconjugating enzymes (E2 or Ubc), and ubiquitin-protein ligases (E3 or Ubr) (7). In an initial activating step the El enzyme forms a thiolester bond with the C terminus of ubiquitin, which is then transferred to a specific cysteine residue of the E2 enzyme. Although E2 enzymes can directly donate ubiquitin to proteins, E3 enzymes are typically required for recognition of specific proteolytic substrates and at least in one case have been shown to be directly involved in the final transfer of ubiquitin to a substrate protein (8). E2 enzymes are encoded by a multigene family and several members of this family have been isolated from various organisms including plants. The function of E2 enzymes has been studied most extensively in yeast by usiing a genetic approach. Mutations in individual genes for dlifferent yeast E2 enzymes revealed that these proteins participate in a wide variety of essential cellular functions (1). In Drosophila an E2 enzyme has been shown to be involved in nervous system development (9, 10). Based on these results it is most likely that in higher organisms, including man, mutations in the genes coding for the different enzymes of the ubiquitin-conljugating system will contribute to abnormal development and disease. Indirect evidence for such a relationship has accumulated over the past years (11).No mouse mutants with spontaneous or experimentally induced alterations have been previously described that help to clarify the role of any of the E2 genes during differentiation and development. In the present report we describe a transgenic mouse mutant where integration of a retrovirus into an E2 gene, designated UbcM4, results in a recessive-lethal phenotype. Homnozygous embryos die in utero most likely as a result of impaired placenta development. MATERIALS AND METHODSDerivation of A6 Mouse Strain. Cells of the embryonal stem (ES) cell line JI (12) were intfected by growing on a monolayer of x-ray irradiated Psi-2 cells producing niplO virus (13) in ES cell culture medium containing 8 ,rg/ml polybrene. After coculture for 48 h, ES cells were trypsinized and selected in medium containing G418 (125 p,g/ml of active component) for 10 days. G418-r...

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Retrovirus-induced insertional mutation in Mov13 mice affects collagen I expression in a tissue-specific manner

Cited by 110 publications

References 40 publications

Restricted expression of Mov13 mutant α1(I) collagen gene in osteoblasts and its consequences for bone development

Restricted expression of Mov13 mutant α1(I) collagen gene in osteoblasts and its consequences for bone development

Mithramycin selectively inhibits collagen-alpha 1(I) gene expression in human fibroblast.

Provirus integration into a gene encoding a ubiquitin-conjugating enzyme results in a placental defect and embryonic lethality.

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