The mRNAs for the pro-alpha 1(I) and pro-alpha 2(I) chains of type I procollagen are translated at the same rate in normal human fibroblasts and in fibroblasts from two variants of osteogenesis imperfecta with altered steady state ratios of the two mRNAs.
Homologous DNA fragments were prepared from cloned cDNAs for the pro-alpha 1(I) and pro-alpha 2(I) chains of human type I procollagen. The DNA fragments were then used to develop a dot blot hybridization assay for mRNAs for pro-alpha 1(I) and pro-alpha 2(I) chains in skin fibroblasts. In normal fibroblasts, the ratio of the steady state levels of the two mRNAs was 1.94 +/- 0.34 S.D. The ratio for the rates of synthesis of the two pro-alpha chains in the same cells was 1.84 +/- 0.13 S.D. Since the two ratios were essentially the same, the results indicated that the mRNAs for the two chains are translated at about the same rates. Therefore, there is no need to invoke translational control or more complex mechanisms to explain synthesis of pro-alpha 1(I) and pro-alpha 2(I) chains in a stoichiometry of 2:1. The dot blot hybridization assay was also used to examine the levels of the mRNAs in fibroblasts from several variants of osteogenesis imperfecta. In two of the variants, the ratios of the steady state levels of mRNAs for pro-alpha 1(I) and pro-alpha 2(I) chains were 3.05 and 2.52, respectively. In the same fibroblasts, the ratios for the rates of synthesis of the two chains were 2.99 +/- 0.43 and 2.45 +/- 0.16, respectively. Therefore, even though the ratios of the levels of the two mRNAs in the fibroblasts were abnormal, the two mRNAs were still translated at the same rates, and there was no evidence of differential regulation at the translational level.
D004247:DNA / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D010013:Osteogenesis Imperfecta / Q000235:genetics*; D011347:Procollagen / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism*
D011347:Procollagen; D012333:RNA, Messenger; D004247:DNA