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Plasma transglutaminase in hypertrophic chondrocytes: expression and cell-specific intracellular activation produce cell death and externalization.

Nurminskaya M, Magee C, Nurminsky D, Linsenmayer TF - J. Cell Biol. (1998)

Bottom Line: We now have isolated a full-length cDNA for this molecule, and confirmed that it is avian factor XIIIA.This externalization most likely is effected by cell death and subsequent lysis-effected by the transglutaminase itself.Non-hypertrophic cells transfected with the same construct do not show these degenerative changes.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Cellular Biology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.

ABSTRACT
We previously used subtractive hybridization to isolate cDNAs for genes upregulated in chick hypertrophic chondrocytes (Nurminskaya, M. , and T.F. Linsenmayer. 1996. Dev. Dyn. 206:260-271). Certain of these showed homology with the "A" subunit of human plasma transglutaminase (factor XIIIA), a member of a family of enzymes that cross-link a variety of intracellular and matrix molecules. We now have isolated a full-length cDNA for this molecule, and confirmed that it is avian factor XIIIA. Northern and enzymatic analyses confirm that the molecule is upregulated in hypertrophic chondrocytes (as much as eightfold). The enzymatic analyses also show that appreciable transglutaminase activity in the hypertrophic zone becomes externalized into the extracellular matrix. This externalization most likely is effected by cell death and subsequent lysis-effected by the transglutaminase itself. When hypertrophic chondrocytes are transfected with a cDNA construct encoding the zymogen of factor XIIIA, the cells convert the translated protein to a lower molecular weight form, and they initiate cell death, become permeable to macromolecules and eventually undergo lysis. Non-hypertrophic cells transfected with the same construct do not show these degenerative changes. These results suggest that hypertrophic chondrocytes have a novel, tissue-specific cascade of mechanisms that upregulate the synthesis of plasma transglutaminase and activate its zymogen. This produces autocatalytic cell death, externalization of the enzyme, and presumably cross-linking of components within the hypertrophic matrix. These changes may in turn regulate the removal and/or calcification of this hypertrophic matrix, which are its ultimate fates.

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Northern analysis of  factor XIIIA expression in hypertrophic (Hyp) and non- hypertrophic (NH) chondrocytes. Total RNA (10 μg) was  isolated from cell cultures  from 14- and 20-d-old embryos. Relative abundance is  normalized to the housekeeping gene GAPDH.
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Figure 1: Northern analysis of factor XIIIA expression in hypertrophic (Hyp) and non- hypertrophic (NH) chondrocytes. Total RNA (10 μg) was isolated from cell cultures from 14- and 20-d-old embryos. Relative abundance is normalized to the housekeeping gene GAPDH.

Mentions: These clones were used as probes for Northern analyses of total RNA from cell cultures of hypertrophic and non-hypertrophic chondrocytes. The data (Fig. 1) give the size of the mRNA as 4.3 kb.


Plasma transglutaminase in hypertrophic chondrocytes: expression and cell-specific intracellular activation produce cell death and externalization.

Nurminskaya M, Magee C, Nurminsky D, Linsenmayer TF - J. Cell Biol. (1998)

Northern analysis of  factor XIIIA expression in hypertrophic (Hyp) and non- hypertrophic (NH) chondrocytes. Total RNA (10 μg) was  isolated from cell cultures  from 14- and 20-d-old embryos. Relative abundance is  normalized to the housekeeping gene GAPDH.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2132883&req=5

Figure 1: Northern analysis of factor XIIIA expression in hypertrophic (Hyp) and non- hypertrophic (NH) chondrocytes. Total RNA (10 μg) was isolated from cell cultures from 14- and 20-d-old embryos. Relative abundance is normalized to the housekeeping gene GAPDH.
Mentions: These clones were used as probes for Northern analyses of total RNA from cell cultures of hypertrophic and non-hypertrophic chondrocytes. The data (Fig. 1) give the size of the mRNA as 4.3 kb.

Bottom Line: We now have isolated a full-length cDNA for this molecule, and confirmed that it is avian factor XIIIA.This externalization most likely is effected by cell death and subsequent lysis-effected by the transglutaminase itself.Non-hypertrophic cells transfected with the same construct do not show these degenerative changes.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Cellular Biology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.

ABSTRACT
We previously used subtractive hybridization to isolate cDNAs for genes upregulated in chick hypertrophic chondrocytes (Nurminskaya, M. , and T.F. Linsenmayer. 1996. Dev. Dyn. 206:260-271). Certain of these showed homology with the "A" subunit of human plasma transglutaminase (factor XIIIA), a member of a family of enzymes that cross-link a variety of intracellular and matrix molecules. We now have isolated a full-length cDNA for this molecule, and confirmed that it is avian factor XIIIA. Northern and enzymatic analyses confirm that the molecule is upregulated in hypertrophic chondrocytes (as much as eightfold). The enzymatic analyses also show that appreciable transglutaminase activity in the hypertrophic zone becomes externalized into the extracellular matrix. This externalization most likely is effected by cell death and subsequent lysis-effected by the transglutaminase itself. When hypertrophic chondrocytes are transfected with a cDNA construct encoding the zymogen of factor XIIIA, the cells convert the translated protein to a lower molecular weight form, and they initiate cell death, become permeable to macromolecules and eventually undergo lysis. Non-hypertrophic cells transfected with the same construct do not show these degenerative changes. These results suggest that hypertrophic chondrocytes have a novel, tissue-specific cascade of mechanisms that upregulate the synthesis of plasma transglutaminase and activate its zymogen. This produces autocatalytic cell death, externalization of the enzyme, and presumably cross-linking of components within the hypertrophic matrix. These changes may in turn regulate the removal and/or calcification of this hypertrophic matrix, which are its ultimate fates.

Show MeSH
Related in: MedlinePlus