title: Functional linkage between the endoplasmic reticulum protein Hsp47 and procollagen expression in human vascular smooth muscle cells.
abstract: Hsp47 is a heat stress protein that interacts with procollagen in the lumen of the endoplasmic reticulum, which is vital for collagen elaboration and embryonic viability. The precise actions of Hsp47 remain unclear, however. To evaluate the effects of Hsp47 on collagen production we infected human vascular smooth muscle cells (SMCs) with a retrovirus containing Hsp47 cDNA. SMCs overexpressing Hsp47 secreted type I procollagen faster than SMCs transduced with empty vector, yielding a greater accumulation of pro alpha1(I) collagen in the extracellular milieu. Interestingly, the amount of intracellular pro alpha1(I) collagen was also increased. This was associated with an unexpected increase in the rate of pro alpha1(I) collagen chain synthesis and 2.5-fold increase in pro alpha1(I) collagen mRNA expression, without a change in fibronectin expression. This amplification of procollagen expression, synthesis, and secretion by Hsp47 imparted SMCs with an enhanced capacity to elaborate a fibrillar collagen network. The effects of Hsp47 were qualitatively distinct from, and independent of, those of ascorbate and the combination of both factors yielded an even more intricate fibril network. Given the in vitro impact of altered Hsp47 expression on procollagen production, we sought evidence for interindividual variability in Hsp47 expression and identified a common, single nucleotide polymorphism in the Hsp47 gene promoter among African Americans that significantly reduced promoter activity. Together, these findings indicate a novel means by which type I collagen production is regulated by the endoplasmic reticulum constituent, Hsp47, and suggest a potential basis for inherent differences in collagen production within the population.
authors.affiliation: Robarts Research Institute, Vascular Biology Group, London Health Sciences Center, Department of Medicine (Cardiology), University of Western Ontario, London N6A 5K8, Canada.
authors.forename: Edward F
authors.initials: EF
authors.lastname: Rocnik
authors.forename: Eric
authors.initials: E
authors.lastname: van der Veer
authors.forename: Henian
authors.initials: H
authors.lastname: Cao
authors.forename: Robert A
authors.initials: RA
authors.lastname: Hegele
authors.forename: J Geoffrey
authors.initials: JG
authors.lastname: Pickering
mesh_terms: D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000737:chemistry* / Q000378:metabolism; D005787:Gene Frequency; D006023:Glycoproteins; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism*; D011347:Procollagen / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
chemical_list: D002352:Carrier Proteins; D024042:Collagen Type I; D006023:Glycoproteins; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D011347:Procollagen; C494660:SERPINH1 protein, human; C046289:colligin