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Global defects in collagen secretion in a Mia3/TANGO1 knockout mouse.

Wilson DG, Phamluong K, Li L, Sun M, Cao TC, Liu PS, Modrusan Z, Sandoval WN, Rangell L, Carano RA, Peterson AS, Solloway MJ - J. Cell Biol. (2011)

Bottom Line: These changes are associated with intracellular accumulation of collagen and the induction of a strong unfolded protein response, primarily within the developing skeleton.Chondrocyte maturation and bone mineralization are severely compromised in Mia3- embryos, leading to dwarfism and neonatal lethality.Thus, Mia3's role in protein secretion is much broader than previously realized, and it may, in fact, be required for the efficient secretion of all collagen molecules in higher organisms.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Biology, Genentech, South San Francisco, CA 94080, USA.

ABSTRACT
Melanoma inhibitory activity member 3 (MIA3/TANGO1) [corrected] is an evolutionarily conserved endoplasmic reticulum resident transmembrane protein. Recent in vitro studies have shown that it is required for the loading of collagen VII, but not collagen I, into COPII-coated transport vesicles. In this paper, we show that mice lacking Mia3 are defective for the secretion of numerous collagens, including collagens I, II, III, IV, VII, and IX, from chondrocytes, fibroblasts, endothelial cells, and mural cells. Collagen deposition by these cell types is abnormal, and extracellular matrix composition is compromised. These changes are associated with intracellular accumulation of collagen and the induction of a strong unfolded protein response, primarily within the developing skeleton. Chondrocyte maturation and bone mineralization are severely compromised in Mia3- embryos, leading to dwarfism and neonatal lethality. Thus, Mia3's role in protein secretion is much broader than previously realized, and it may, in fact, be required for the efficient secretion of all collagen molecules in higher organisms.

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The UPR is up-regulated in chondrocytes of Mia3- animals coincident with delayed bone maturation. (A) Gene expression in wt and Mia3- 14.5-dpc limbs (n = 5 each) was studied by Mouse Genome v2.0 arrays (Affymetrix) with probes for over 39,000 transcripts. Using unsupervised gene ontology analysis (Broad Institute, Cambridge, MA), over-represented patterns of genes associated with ER and Golgi function/UPR/ERAD were identified. All data presented have a P < 0.05. (B and C) Comparison of array data for several probes specific to chondrogenic/osteogenic progression (B; Col10a1, Dmp1, Ibsp, and Spp1) and UPR/ER stress (C; Atf4, Atf5, Ddit3, Chac1, Eif4ebp1, Creb3, Hspa5, and Slc7a3) in 12.5-, 13.5-, and 14.5-dpc whole forelimbs. Misregulation of both gene sets in mutants is concomitant at 13.5 dpc. Col10a1 and the UPR gene set are significantly misregulated (P < 0.05) at 13.5 dpc, whereas the osteogenic markers Dmp1, Ibsp, and Spp1 achieve significance at 14.5 dpc. GEO Datasets accession numbers are given at the top. (D–M′) Section in situ hybridization of 14.5-dpc humeri from wt and Mia3- embryos using probes against Atf4, Atf5, Ddit3, Chac1, and Trib3 reveals prominent staining throughout all chondrocytes in comparison to controls. D′–M′ denotes close-ups from the humeri shown in D–M. Bars, 100 µm.
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fig8: The UPR is up-regulated in chondrocytes of Mia3- animals coincident with delayed bone maturation. (A) Gene expression in wt and Mia3- 14.5-dpc limbs (n = 5 each) was studied by Mouse Genome v2.0 arrays (Affymetrix) with probes for over 39,000 transcripts. Using unsupervised gene ontology analysis (Broad Institute, Cambridge, MA), over-represented patterns of genes associated with ER and Golgi function/UPR/ERAD were identified. All data presented have a P < 0.05. (B and C) Comparison of array data for several probes specific to chondrogenic/osteogenic progression (B; Col10a1, Dmp1, Ibsp, and Spp1) and UPR/ER stress (C; Atf4, Atf5, Ddit3, Chac1, Eif4ebp1, Creb3, Hspa5, and Slc7a3) in 12.5-, 13.5-, and 14.5-dpc whole forelimbs. Misregulation of both gene sets in mutants is concomitant at 13.5 dpc. Col10a1 and the UPR gene set are significantly misregulated (P < 0.05) at 13.5 dpc, whereas the osteogenic markers Dmp1, Ibsp, and Spp1 achieve significance at 14.5 dpc. GEO Datasets accession numbers are given at the top. (D–M′) Section in situ hybridization of 14.5-dpc humeri from wt and Mia3- embryos using probes against Atf4, Atf5, Ddit3, Chac1, and Trib3 reveals prominent staining throughout all chondrocytes in comparison to controls. D′–M′ denotes close-ups from the humeri shown in D–M. Bars, 100 µm.

Mentions: Consistent with these morphological changes, gene expression profiling by the microarray of wt and Mia3−/− forelimb RNA collected from 12.5-, 13.5-, and 14.5-dpc embryos (n = 5 per genotype) reveals a highly significant increase in genes associated with the UPR (Fig. 8 A). These include primary drivers of the UPR, such as CHOP and ATF4, as well as numerous downstream targets, including Grp78/BiP (binding immunoglobulin protein), Atf5, and Dnajb9. Mediators of ER-associated degradation (ERAD), such as Derl1 and Edem1, are also highly up-regulated. The majority of these genes are expressed at normal (low) levels at 12.5 dpc, but their expression begins to diverge from wt littermates at 13.5 dpc (Fig. 8, B and C). This trend was maintained at 14.5 and 16.5 dpc (Fig. S5).


Global defects in collagen secretion in a Mia3/TANGO1 knockout mouse.

Wilson DG, Phamluong K, Li L, Sun M, Cao TC, Liu PS, Modrusan Z, Sandoval WN, Rangell L, Carano RA, Peterson AS, Solloway MJ - J. Cell Biol. (2011)

The UPR is up-regulated in chondrocytes of Mia3- animals coincident with delayed bone maturation. (A) Gene expression in wt and Mia3- 14.5-dpc limbs (n = 5 each) was studied by Mouse Genome v2.0 arrays (Affymetrix) with probes for over 39,000 transcripts. Using unsupervised gene ontology analysis (Broad Institute, Cambridge, MA), over-represented patterns of genes associated with ER and Golgi function/UPR/ERAD were identified. All data presented have a P < 0.05. (B and C) Comparison of array data for several probes specific to chondrogenic/osteogenic progression (B; Col10a1, Dmp1, Ibsp, and Spp1) and UPR/ER stress (C; Atf4, Atf5, Ddit3, Chac1, Eif4ebp1, Creb3, Hspa5, and Slc7a3) in 12.5-, 13.5-, and 14.5-dpc whole forelimbs. Misregulation of both gene sets in mutants is concomitant at 13.5 dpc. Col10a1 and the UPR gene set are significantly misregulated (P < 0.05) at 13.5 dpc, whereas the osteogenic markers Dmp1, Ibsp, and Spp1 achieve significance at 14.5 dpc. GEO Datasets accession numbers are given at the top. (D–M′) Section in situ hybridization of 14.5-dpc humeri from wt and Mia3- embryos using probes against Atf4, Atf5, Ddit3, Chac1, and Trib3 reveals prominent staining throughout all chondrocytes in comparison to controls. D′–M′ denotes close-ups from the humeri shown in D–M. Bars, 100 µm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3105544&req=5

fig8: The UPR is up-regulated in chondrocytes of Mia3- animals coincident with delayed bone maturation. (A) Gene expression in wt and Mia3- 14.5-dpc limbs (n = 5 each) was studied by Mouse Genome v2.0 arrays (Affymetrix) with probes for over 39,000 transcripts. Using unsupervised gene ontology analysis (Broad Institute, Cambridge, MA), over-represented patterns of genes associated with ER and Golgi function/UPR/ERAD were identified. All data presented have a P < 0.05. (B and C) Comparison of array data for several probes specific to chondrogenic/osteogenic progression (B; Col10a1, Dmp1, Ibsp, and Spp1) and UPR/ER stress (C; Atf4, Atf5, Ddit3, Chac1, Eif4ebp1, Creb3, Hspa5, and Slc7a3) in 12.5-, 13.5-, and 14.5-dpc whole forelimbs. Misregulation of both gene sets in mutants is concomitant at 13.5 dpc. Col10a1 and the UPR gene set are significantly misregulated (P < 0.05) at 13.5 dpc, whereas the osteogenic markers Dmp1, Ibsp, and Spp1 achieve significance at 14.5 dpc. GEO Datasets accession numbers are given at the top. (D–M′) Section in situ hybridization of 14.5-dpc humeri from wt and Mia3- embryos using probes against Atf4, Atf5, Ddit3, Chac1, and Trib3 reveals prominent staining throughout all chondrocytes in comparison to controls. D′–M′ denotes close-ups from the humeri shown in D–M. Bars, 100 µm.
Mentions: Consistent with these morphological changes, gene expression profiling by the microarray of wt and Mia3−/− forelimb RNA collected from 12.5-, 13.5-, and 14.5-dpc embryos (n = 5 per genotype) reveals a highly significant increase in genes associated with the UPR (Fig. 8 A). These include primary drivers of the UPR, such as CHOP and ATF4, as well as numerous downstream targets, including Grp78/BiP (binding immunoglobulin protein), Atf5, and Dnajb9. Mediators of ER-associated degradation (ERAD), such as Derl1 and Edem1, are also highly up-regulated. The majority of these genes are expressed at normal (low) levels at 12.5 dpc, but their expression begins to diverge from wt littermates at 13.5 dpc (Fig. 8, B and C). This trend was maintained at 14.5 and 16.5 dpc (Fig. S5).

Bottom Line: These changes are associated with intracellular accumulation of collagen and the induction of a strong unfolded protein response, primarily within the developing skeleton.Chondrocyte maturation and bone mineralization are severely compromised in Mia3- embryos, leading to dwarfism and neonatal lethality.Thus, Mia3's role in protein secretion is much broader than previously realized, and it may, in fact, be required for the efficient secretion of all collagen molecules in higher organisms.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Biology, Genentech, South San Francisco, CA 94080, USA.

ABSTRACT
Melanoma inhibitory activity member 3 (MIA3/TANGO1) [corrected] is an evolutionarily conserved endoplasmic reticulum resident transmembrane protein. Recent in vitro studies have shown that it is required for the loading of collagen VII, but not collagen I, into COPII-coated transport vesicles. In this paper, we show that mice lacking Mia3 are defective for the secretion of numerous collagens, including collagens I, II, III, IV, VII, and IX, from chondrocytes, fibroblasts, endothelial cells, and mural cells. Collagen deposition by these cell types is abnormal, and extracellular matrix composition is compromised. These changes are associated with intracellular accumulation of collagen and the induction of a strong unfolded protein response, primarily within the developing skeleton. Chondrocyte maturation and bone mineralization are severely compromised in Mia3- embryos, leading to dwarfism and neonatal lethality. Thus, Mia3's role in protein secretion is much broader than previously realized, and it may, in fact, be required for the efficient secretion of all collagen molecules in higher organisms.

Show MeSH
Related in: MedlinePlus