Construction and characterization of type II collagen complementary deoxyribonucleic acid dones

Lukens, Lewis; Frischauf, Anna Maria; Pawlowski, Philip J.; Brierley, George T.; Lehrach, Hans

doi:10.1093/nar/11.17.6021

Cited by 23 publications

(17 citation statements)

References 29 publications

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“…1). A very prominent RNA of 5,100 bases was detected in normal chondrocytes, as described previously (16,28,57). Transformed chondrocytes displayed reductions in this RNA species to 1.5% (wt-RSV-transformed cells) and 3% (ts-RSVtransformed cells) of normal levels, consistent with the dramatic reduction in the rate of type II collagen synthesis in these cells (2).…”

supporting

confidence: 75%

“…The type II collagen gene does not possess an inverted repeat sequence in the translation initiation region (21) and does not possess multiple consensus signals for polyadenylation (35,44). Furthermore, although there is one report of multiple type II collagen RNAs (57), recent reports from other laboratories have not shown this (16,21,28,65).…”

mentioning

confidence: 96%

“…The following probes were used in the Northern hybridizations: pFN600, a 600-base-pair fibronectin cDNA (14); pCOL3, an 800-base-pair al(I) collagen cDNA (64); pCg45, a 2,500-base-pair a2(I) collagen cDNA (24); pCgII-12, a 740-base-pair al(II) collagen cDNA (28); and pHrB, a human 18S ribosomal DNA (59). The plasmids pFN600, pCg45, pCgII-12, and pHrB were generous gifts of K. Yamada RNAs from normal and transformed chondrocytes were subjected to Northern hybridization analysis with 32P-labeled pCgII-12, a cloned type II collagen cDNA (28) (Fig.…”

mentioning

confidence: 99%

“…The plasmids pFN600, pCg45, pCgII-12, and pHrB were generous gifts of K. Yamada RNAs from normal and transformed chondrocytes were subjected to Northern hybridization analysis with 32P-labeled pCgII-12, a cloned type II collagen cDNA (28) (Fig. 1).…”

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confidence: 99%

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Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation.

Allebach¹,

Boettiger²,

Pacifici³

et al. 1985

Mol. Cell. Biol.

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We have analyzed the effects of transformation by Rous sarcoma virus on expression of types I and II collagen and fibronectin genes in vertebral chondrocytes and compared them with expression of these genes in skin fibroblasts. Transformed chondrocytes display a dramatically decreased amount of type II collagen RNA, which can account fully for the decreased synthetic rate of this protein. Paradoxically, these cells also display greatly increased amounts of type I collagen RNAs, which are translated efficientlv in vitro, but not in the intact cells. We show here that the type I collagen RNAs in transformed chondrocytes are nearly indistinguishable from those found in skin fibroblasts, and that they clearly differ from the type I collagen RNAs found in normal chondrocytes. Transformed chondrocytes also display an increased amount of fibronectin RNA, which can account fully for the increased synthetic rate of this protein. Thus, the effects of transformation by Rous sarcoma virus on type I collagen and fibronectin RNAs in chondrocytes are the opposite of those observed in fibroblasts, which display decreased amounts of these three RNAs. These data indicate that the effects of transformation on the genes encoding type I collagen and fibronectin must be modulated by host cell-specific factors. They also imply that the types I and II collagen genes may be regulated by different mechanisms, the type I genes being controlled at both transcriptional and posttranscriptional levels, and the type II gene being controlled primarily at the transcriptional level.Transformation of differentiated cells by tumor viruses has provided a useful tool for analyzing the mechanisms of cellular gene expression. This has been particularly true for the genes encoding extracellular matrix proteins, whose expression is decreased by transformation with certain viruses. For example, transformation of chicken embryo fibroblasts by Rous sarcoma virus (RSV) results in reduced synthesis of type I collagen (10,20,25,(45)(46)(47) and, in some cases, fibronectin (36,39). Similarly, transformation of chicken vertebral chondrocytes by RSV results in decreased synthesis of type II collagen and the major cartilage proteoglycan (2,18,37,38). The reduced rates of synthesis of type I collagen and fibronectin in transformed fibroblasts reflect decreased amounts of the corresponding RNAs (1, 3. 15,19,43,45,46,51) These data demonstrate that transformation by RSV has opposite effects on the expression of type I collagen and fibronectin genes in chondrocytes and fibroblasts and imply that the action of src is modulated by host cell factors that differ between these cell types. They also imply that the types I and II collagen genes may be regulated by different mechanisms. The genes encoding type I collagen, a protein which is synthesized by many cell types, appear to be regulated at both transcriptional and posttranscriptional levels. In contrast, the type II collagen gene, which is expressed only by a very limited number of terminally differentiated cell type...

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supporting

confidence: 75%

mentioning

confidence: 96%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation.

Allebach¹,

Boettiger²,

Pacifici³

et al. 1985

Mol. Cell. Biol.

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“…Gerstenfeld et al (18) have shown that quantitative separation of RNA from DNA can be achieved by using this modification of the original GuHCl RNA isolation procedure (12). We found this method to give greater yields of intact mRNA despite previous reports which suggested that the GuSCN method (12) was more efficient for extraction of RNA from chondrocytes (31). Polyadenylated mRNA was prepared by the method of Tate et al (47).…”

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confidence: 79%

Collagen expression in embryonic chicken chondrocytes treated with phorbol myristate acetate.

Finer¹,

Gerstenfeld²,

Young³

et al. 1985

Mol. Cell. Biol.

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Growth of embryonic chicken sternal chondrocytes in the presence of phorbol-12-myristate-13-acetate (PMA), a potent tumor promoter, resulted in a dramatic morphological change from spherical floating cells to adherent fibroblastic cells. This morphological change was accompanied by a quantitative switch from synthesis of cartilage-specific type II procollagen to type I procollagen. Type II procollagen mRNA levels decreased 10-fold in PMA-treated cells. Activation of type I collagen genes led to the accumulation of type I procollagen mRNA levels comparable to those of type H mRNA in these cells. However, only type I procollagen mRNA was translated. In addition to gene activation, unprocessed pro al(l) transcripts present at low levels in control chondrocytes were processed to mature mRNA species. Redifferentiation of PMA-treated chondrocytes was possible if cells were removed from PMA after the morphological change and cessation of type H procollagen synthesis but before detectable amounts of type I procollagen were synthesized. Production of type I collagen thus marks a late phase of chondrocyte "dedifferentiation" from which reversion is no longer possible. Redifferentiated cell populations contained 24-fold more pro al(II) collagen mRNA than pro al(I) collagen mRNA, but the rates of procollagen synthesis were comparable. This suggests that the PMA-mediated dedifferentiation of chondrocytes as well as their redifferentiation is under both transcriptional and posttranscriptional regulation.In vivo and in vitro, differentiated chondrocytes secrete and are imbedded in an extracellular matrix composed of type II collagen (16,36,44,50), several minor collagens specific to hyaline cartilage (33), and chondrocyte-specific type IV sulfated proteoglycans (14,20). The latter can be identified because they stain with toluidine blue (13). Chondrocytes in culture are either spherical and remain in suspension or attach to the culture dish and assume a characteristic polygonal morphology (50). When subcultured in monolayer, the cells gradually assume a more fibroblast-like morphology. Morphological changes are followed by a decrease in the expression of the chondrocyte matrix components, including type II collagen and type IV sulfated proteoglycans. Concomitantly, low levels of type I collagen and fibronectin, characteristic of the extracellular matrix of fibroblasts in skin, bone, and tendons are produced. These changes are not the result of overgrowth by contaminating fibroblasts since the switch from type II to type I collagen production can be observed in the progeny of a cloned chondrocyte (10,15,34). The failure to continue to produce the differentiated phenotype led to the characterization of this switch as "dedifferentiation" (22). This is not meant to imply that the cells had returned to an uncommitted or multipotent state, but rather that adverse environmental conditions have interrupted the expression of the differentiated phenotype, somewhat like the heat shock effect. As a result, the cells either revert to an ea...

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Loss of a tumor suppressor gene function is correlated with downregulation of chondrocyte‐specific collagen expression in syrian hamster embryo cells

Hosoi

Montgomery

et al. 1991

Molecular Carcinogenesis

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We previously described the isolation of closely related, preneoplastic Syrian hamster cell lines that have retained (supB+) or lost (supB-) the ability to suppress the anchorage-independent growth and tumorigenicity of a sarcoma cell line (BP6T) in cell hybrids. In this report, we have used differential cDNA screening to clone several genes that are expressed in supB+ cells and downregulated in supB- cells. The nontumorigenic supB+ and supB- variants are advantageous for differential cDNA cloning because multiple independent cell lines differing in their tumor suppressor activity have been isolated. Differentially expressed cDNAs were isolated and placed into one of four groups based on DNA cross-hybridization. Representative cDNAs from Groups I and II, which were expressed at relatively high levels in two independently derived supB+ cell lines (DES4 and 10W) and downregulated in the supB- and tumor cell lines, were sequenced. The DNA and predicted amino acid sequences of these genes were found to be highly homologous to the chondrocyte-specific collagens type II and type IX. In contrast to the chondrocyte-specific collagens, another collagen isoform, collagen type I, was expressed at similar levels in both supB+ and supB- cells. These results suggest that carcinogen-induced immortalization selected for chondrocyte-like cell lines from the mixed embryo cell population. As these cells progressed toward tumorigenicity, the ability to express the chondrocyte differentiation markers was lost concomitantly with the ability to suppress the tumorigenicity of the BP6T sarcoma cell line. These results are consistent with the hypothesis that the supB+ tumor suppressor gene is involved in the regulation of differentiation. The identification of genes regulated by this suppressor gene may aid in its isolation.

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Construction and characterization of type II collagen complementary deoxyribonucleic acid dones

Cited by 23 publications

References 29 publications

Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation.

Control of types I and II collagen and fibronectin gene expression in chondrocytes delineated by viral transformation.

Collagen expression in embryonic chicken chondrocytes treated with phorbol myristate acetate.

Loss of a tumor suppressor gene function is correlated with downregulation of chondrocyte‐specific collagen expression in syrian hamster embryo cells

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