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Cloning and Expression Analysis of the ${\alpha}$-Subunit of Porcine Prolyl 4-hydroxylase

  • Cho, Eun Seok (Department of Animal Resources Technology, Jinju National University) ;
  • Jung, Won Youg (Department of Animal Resources Technology, Jinju National University) ;
  • Kwon, Eun Jung (Department of Animal Resources Technology, Jinju National University) ;
  • Park, Da Hye (Department of Animal Resources Technology, Jinju National University) ;
  • Chung, Ki Hwa (Department of Animal Resources Technology, Jinju National University) ;
  • Cho, Kwang Keun (Department of Animal Resources Technology, Jinju National University) ;
  • Kim, Chul Wook (Department of Animal Resources Technology, Jinju National University)
  • 투고 : 2007.03.05
  • 심사 : 2007.06.05
  • 발행 : 2007.11.01

초록

Prolyl 4-hydroxylase (P4H) plays a central role in collagen synthesis by catalyzing the hydroxylation of the proline residue in the X-Pro-Gly amino acid sequence, and controls the biosynthesis of collagen that influences overall meat quality. In order to verify expression level of the catalytic ${\alpha}$ subunit of P4H, a 2.7 kb clone of the ${\alpha}$ subunit gene for P4H was selected from a cDNA library prepared from the muscular tissue of Sancheong berkshire pigs, and the whole gene sequence was determined. As expression level of the ${\alpha}$ subunit of P4H differed between tissues of pigs, we intended to assess more precisely the level of ${\alpha}$-subunit expression between tissues of Sancheong Berkshire pigs by using RT-PCR. Muscular and adipose tissues were taken from each pig grouped by growth stage (weighing 60, 80, and 110 kg) of Yorkshire and Sancheong Berkshire pigs, and the expression levels of the ${\alpha}$-subunit of P4H were examined. Since there were significant differences in the expression level with respect to variation in growth stage (p<0.01), an attempt was made to identify any influences of pig species and tissue variation. The muscular and adipose tissues of pigs weighing 110 kg showed higher expression levels than pigs weighing 60 kg and 80 kg. In general, significantly higher expression levels were found in muscular than in adipose tissue. The expression levels in Sancheong Berkshire were significantly higher than in Yorkshire pigs (p<0.01 or p<0.05). Since expression level of the ${\alpha}$-subunit of P4H affects the activity of P4H and is connected to the biosynthesis of collagen and increased collagen can improve meat texture, this finding may explain why meat quality of the Sancheong Berkshire pig is acclaimed in Korea. Given the higher expression levels of the ${\alpha}$-subunit gene in adipose than in muscular tissue, and also in the heavier pigs, more intensive studies are required to assess the correlation between expression level of the ${\alpha}$ subunit gene and overall meat quality.

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참고문헌

  1. Bassuk, J. A., W. W.-Y. Kao, P. Herzer, N. L. Kedersha, J. Seyer, J. A. DeMartino, B. L. Daugherty, G. E. Mark and R. A. Berg. 1989. Prolyl hydroxylase: molecular cloning and primary structure of the ${\alpha}$ subunit from chicken embryo. Proc. Natl. Acad. Sci. USA. 86:7382-7386. https://doi.org/10.1073/pnas.86.19.7382
  2. Bassuk, J. A. and R. A. Berg. 1991. Correlation of the steady-state RNA levels among the ${\alpha}1$ and ${\alpha}2$ chains of type 1 collagen during growth of chicken embryo tendon fibroblasts. Biochem. Biophys. Res. Commun. 174:169-175. https://doi.org/10.1016/0006-291X(91)90501-W
  3. Han, X-Y., W. Wang, R. Myllyla, P. Virtanen, J. Karpakka and T. E. S. Takala. 1999. mRNA levels for ${\alpha}-subunit$ of prolyl 4-hydroxylase and fibrillar collagens in immobilized rat skeletal muscle. J. Appl. Physiol. 87(1):90-96. https://doi.org/10.1152/jappl.1999.87.1.90
  4. Helaakoski, T., K. Vuori, R. Myllyla, K. I. Kivirikko and T. Pihlajaiemi. 1989. Molecular cloning of the ${\alpha}-subunit$ of human prolyl 4-hydroxylase: the complete cDNA-derived amino acid sequence and evidence for alternative splicing of RNA transcripts. Proc. Natl. Acad. Sci. USA. 86:4392-4396. https://doi.org/10.1073/pnas.86.12.4392
  5. Helaakoski, T., J. Veijola, K. Vuori, M. Rehn, L. T. Chow, P. Taillon-Miller, K. I. Kivirikko and T. Pihlajaniemi. 1994. Structure and Expression of the human gene for the ${\alpha}\;subunit$ of prolyl 4-hydroxylase. J. Biological Chem. 269(45):27847-27854.
  6. Johnstone, I. L. 2000. Cuticle collagen genes. Expression in Caenorhabditiselegans. Trends. Genet. 16:21-27.
  7. JOrgensen, F. G., A. Hobolth, HornshOj, C. Bendixen, M. Fredholm and M. H. Schierup. 2005. Comparative analysis of protein coding sequences from human, mouse and the domesticated pig. BMC Biol. 3:2. https://doi.org/10.1186/1741-7007-3-2
  8. Kao, W. W. Y., C. W.-C. Kao and R. I. Schwarz. 1985. Prolyl hydroxylase production can be uncoupled from the regulation of procollagen synthesis. Exp. Cell Res. 157:265-270. https://doi.org/10.1016/0014-4827(85)90168-5
  9. Kim, C. W., K. T. Chang, Y. H. Hong, E. J. Kwon, W. Y. Jung, K. K. Cho, K. H. Chung, B. W. Kim, J. G. Lee, J. S. Yeo, Y. S. Kang and Y. K. Joo. 2005. Screening of specific genes expressed in the swine tissues and development of a functional cDNA chip. Asian-Aust. J. Anim. Sci. 18(7):933-941. https://doi.org/10.5713/ajas.2005.933
  10. Kivirikko, K. I., R. Myllyla and T. Pihlajaniemi. 1992. Hydroxylation of praline and lysine residues in collagens and other animal and plant proteins. In: (Ed. J. J. Harding and J. C. Crabbe) Post-Translational Modifications of Protein, CRC Press, Boca Raton, FL, pp. 1-51.
  11. Kivirikko, K. I. and T. Pihlajaniemi. 1998. Collagen hydroxylases and the protein disulfide isomerase subunit of prolyl 4-hydroxylases. Adv. Enzymol. Related Areas Mol. Biol. 72:325-398.
  12. Kivirikko, K. I. and J. Myllharju. 1998. Prolyl 4-hydroxylases and their protein disulfide isomerase subunit. Matrix Biol. 16:357-368. https://doi.org/10.1016/S0945-053X(98)90009-9
  13. Lee, E. H., W. W.-Y. Kao and R. I. Schwarz. 2001. Cell density regulates prolyl 4-hydroxylase activity independent of mRNA levels. Matrix Biol. 19:779-782. https://doi.org/10.1016/S0945-053X(00)00123-2
  14. Mayne, R. 1982. Collagen in health and disease. New York:Churchill Livingstone, 445-455.
  15. Mendel, M. and A. Higa. 1970. Calcium dependent bacteriophase DNA detection. J. Molec. Biol. 53:159-162. https://doi.org/10.1016/0022-2836(70)90051-3
  16. Myllyharju, J. 2003. Prolyl 4-hydroylases, the key enzymes of collagen biosynthesis. Matrix Biol. 22:15-24. https://doi.org/10.1016/S0945-053X(03)00006-4
  17. Rowe, L. B. and R. I. Schwarz. 1983. Role of procollagen mRNA levels in controlling the rate of procollagen synthesis. Mol. Cell. Biol. 3:241-249. https://doi.org/10.1128/MCB.3.2.241
  18. Tadayosi, N. 2003. Organisms in Natural Features; Production of a native black pig . Gyeongsangnam-do, pp. 11-21.

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