Structure of the gene encoding the beta-subunit of chicken prolyl 4-hydroxylase

• Published in: Gene Vol. 71; no. 2; p. 451
• Main Authors: Nakazawa, M, Aida, T, Everson, W V, Gonda, M A, Hughes, S H, Kao, W W
• Format: Journal Article
• Language: English
• Published: Netherlands 30-11-1988
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Chickens; DNA - genetics; Endonucleases - antagonists & inhibitors; Exons; Genes; Introns; Molecular Sequence Data; Nucleic Acid Heteroduplexes - analysis; Procollagen-Proline Dioxygenase - genetics; Promoter Regions, Genetic; Restriction Mapping; Single-Strand Specific DNA and RNA Endonucleases; Transcription, Genetic
• ISSN: 0378-1119
• DOI: 10.1016/0378-1119(88)90062-5

Prolyl 4-hydroxylase (EC 1.14.11.2) converts peptidyl proline to peptidyl hydroxyproline in procollagen polypeptides during collagen biosynthesis. The active enzyme is a tetramer which is composed of two pairs of non-identical subunits in a molecular form of alpha 2 beta 2. In addition to the tetrameric prolyl 4-hydroxylase (alpha 2 beta 2), the free beta-subunit is also found inside cells. Recently it was shown that the beta-subunit of prolyl 4-hydroxylase is identical to the protein disulfide isomerase and cellular thyroid hormone-binding protein. We previously isolated and characterized cDNAs of the beta-subunit of chicken prolyl 4-hydroxylase. The cDNA of beta-subunit was used to screen a chicken genomic DNA library constructed with the lambda EMBL-3 vector. Two clones, lambda gCPH beta-22 and beta-50, were isolated and characterized by restriction enzyme analysis, heteroduplex analysis, and nucleotide sequencing. The results showed that the 2.5-kb mRNA of the beta-subunit is divided into eleven exons and that the gene is 9.0 kb long. The gene contains consensus sequence for TATA at -24 bp and four CAAT at -57, -157, -194 and -223 bp in the 5' end flanking sequence of the transcription start point. In addition, there are three GC boxes upstream from the TATA box and four GC boxes in the first intron. This is similar to the structure of the alpha 1(I) collagen coding gene (COL1A1). These elements may interact with nuclear factors and play important roles in expression regulation of the beta-subunit gene as has been described in COL1A1.