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. 1994 Apr;69(4):738–742. doi: 10.1038/bjc.1994.139

Protein kinase A (PK-A) regulatory subunit expression in colorectal cancer and related mucosa.

A W Bradbury 1, D C Carter 1, W R Miller 1, Y S Cho-Chung 1, T Clair 1
PMCID: PMC1968829  PMID: 8142263

Abstract

Photoaffinity labelling (PAL) with [32P]8-azido-cAMP and polyacrylamide gel electrophoresis (PAGE) has been used to identify three specific cAMP-binding proteins (cAMP-BPs) within cytosols derived from the centre and periphery of 32 human colorectal cancers and from related adjacent (less than 5 cm from the tumour) and distant (more than 5 cm from the tumour) microscopically benign mucosa. By immunoprecipitation with specific anti-RI and anti-RII antibodies these proteins have subsequently been characterised as a single form of RI (48 kDa) and two forms of RII (50 and 52 kDa). The relative expression of isoforms in each specimen has been quantified by laser densitometry. There was significantly more RI expressed in both tumour centre and periphery than in either adjacent or distant mucosa (P < 0.008 by Wilcoxon signed-rank test). There was no significant difference in relative RI expression between tumour centre and periphery, or between adjacent and distant mucosa. There was no association between relative RI expression and Dukes' stage. Poorly differentiated tumours expressed significantly more RI than those that were either moderately or well differentiated (P = 0.016 by Mann-Whitney U-test). This study is the first to have characterised cAMP-BPs within human colorectal tissues and has demonstrated that colorectal cancers, and in particular those of poor histological grade, relatively overexpress RI when compared with related benign mucosa.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ball R. L., Tanner K. D., Carpenter G. Epidermal growth factor potentiates cyclic AMP accumulation in A-431 cells. J Biol Chem. 1990 Aug 5;265(22):12836–12845. [PubMed] [Google Scholar]
  2. Bradbury A. W., Miller W. R., Carter D. C. Cyclic adenosine 3',5'-monophosphate binding proteins in human colorectal cancer and mucosa. Br J Cancer. 1991 Feb;63(2):201–204. doi: 10.1038/bjc.1991.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Cho-Chung Y. S. Site-selective 8-chloro-cyclic adenosine 3',5'-monophosphate as a biologic modulator of cancer: restoration of normal control mechanisms. J Natl Cancer Inst. 1989 Jul 5;81(13):982–987. doi: 10.1093/jnci/81.13.982. [DOI] [PubMed] [Google Scholar]
  5. De Camilli P., Moretti M., Donini S. D., Walter U., Lohmann S. M. Heterogeneous distribution of the cAMP receptor protein RII in the nervous system: evidence for its intracellular accumulation on microtubules, microtubule-organizing centers, and in the area of the Golgi complex. J Cell Biol. 1986 Jul;103(1):189–203. doi: 10.1083/jcb.103.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gentleman S., Hemmings B. A., Russell P., Chader G. J. Developmental expression of the RI subunit of cyclic AMP-dependent protein kinase in retina. Exp Eye Res. 1989 Jun;48(6):717–731. doi: 10.1016/0014-4835(89)90059-6. [DOI] [PubMed] [Google Scholar]
  7. Gentleman S., Russell P., Hemmings B. A., Chader G. J. Abnormal expression of the RI subunit of cyclic AMP-dependent protein kinase in Y-79 retinoblastoma cells. Exp Eye Res. 1989 Apr;48(4):497–507. doi: 10.1016/0014-4835(89)90033-x. [DOI] [PubMed] [Google Scholar]
  8. Gharrett A. J., Malkinson A. M., Sheppard J. R. Cyclic AMP-dependent protein kinases from normal and SV40-transformed 3T3 cells. Nature. 1976 Dec 16;264(5587):673–675. doi: 10.1038/264673a0. [DOI] [PubMed] [Google Scholar]
  9. Haddox M. K., Roeske W. R., Russell D. H. Independent expression of cardiac type I and II cyclic AMP-dependent protein kinase during murine embryogenesis and postnatal development. Biochim Biophys Acta. 1979 Jul 18;585(4):527–534. doi: 10.1016/0304-4165(79)90185-5. [DOI] [PubMed] [Google Scholar]
  10. Jahnsen T., Hedin L., Kidd V. J., Beattie W. G., Lohmann S. M., Walter U., Durica J., Schulz T. Z., Schiltz E., Browner M. Molecular cloning, cDNA structure, and regulation of the regulatory subunit of type II cAMP-dependent protein kinase from rat ovarian granulosa cells. J Biol Chem. 1986 Sep 15;261(26):12352–12361. [PubMed] [Google Scholar]
  11. Jahnsen T., Hedin L., Lohmann S. M., Walter U., Richards J. S. The neural type II regulatory subunit of cAMP-dependent protein kinase is present and regulated by hormones in the rat ovary. J Biol Chem. 1986 May 25;261(15):6637–6639. [PubMed] [Google Scholar]
  12. Lee D. C., Carmichael D. F., Krebs E. G., McKnight G. S. Isolation of a cDNA clone for the type I regulatory subunit of bovine cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3608–3612. doi: 10.1073/pnas.80.12.3608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Linask K. K., Greene R. M. Subcellular compartmentalization of cAMP-dependent protein kinase regulatory subunits during palate ontogeny. Life Sci. 1989;45(20):1863–1868. doi: 10.1016/0024-3205(89)90539-0. [DOI] [PubMed] [Google Scholar]
  14. Lorimer I. A., Mason M. E., Sanwal B. D. Levels of type I cAMP-dependent protein kinase regulatory subunit are regulated by changes in turnover rate during skeletal myogenesis. J Biol Chem. 1987 Dec 15;262(35):17200–17205. [PubMed] [Google Scholar]
  15. Oyen O., Myklebust F., Scott J. D., Hansson V., Jahnsen T. Human testis cDNA for the regulatory subunit RII alpha of cAMP-dependent protein kinase encodes an alternate amino-terminal region. FEBS Lett. 1989 Mar 27;246(1-2):57–64. doi: 10.1016/0014-5793(89)80253-4. [DOI] [PubMed] [Google Scholar]
  16. Pomerantz A. H., Rudolph S. A., Haley B. E., Greengard P. Photoaffinity labeling of a protein kinase from bovine brain with 8-azidoadenosine 3',5'-monophosphate. Biochemistry. 1975 Aug 26;14(17):3858–3862. doi: 10.1021/bi00688a019. [DOI] [PubMed] [Google Scholar]
  17. Rangel-Aldao R., Kupiec J. W., Rosen O. M. Resolution of the phosphorylated and dephosphorylated cAMP-binding proteins of bovine cardiac muscle by affinity labeling and two-dimensional electrophoresis. J Biol Chem. 1979 Apr 10;254(7):2499–2508. [PubMed] [Google Scholar]
  18. Rannels S. R., Corbin J. D. Characterization of small cAMP-binding fragments of cAMP-dependent protein kinases. J Biol Chem. 1979 Sep 10;254(17):8605–8610. [PubMed] [Google Scholar]
  19. Sandberg M., Levy F. O., Oyen O., Hansson V., Jahnsen T. Molecular cloning, cDNA structure and deduced amino acid sequence for the hormone-induced regulatory subunit (RII beta) of cAMP-dependent protein kinase from rat ovarian granulosa cells. Biochem Biophys Res Commun. 1988 Jul 29;154(2):705–711. doi: 10.1016/0006-291x(88)90197-0. [DOI] [PubMed] [Google Scholar]
  20. Sandberg M., Skålhegg B., Jahnsen T. The two mRNA forms for the type I alpha regulatory subunit of cAMP-dependent protein kinase from human testis are due to the use of different polyadenylation site signals. Biochem Biophys Res Commun. 1990 Feb 28;167(1):323–330. doi: 10.1016/0006-291x(90)91768-n. [DOI] [PubMed] [Google Scholar]
  21. Scott J. D., Glaccum M. B., Zoller M. J., Uhler M. D., Helfman D. M., McKnight G. S., Krebs E. G. The molecular cloning of a type II regulatory subunit of the cAMP-dependent protein kinase from rat skeletal muscle and mouse brain. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5192–5196. doi: 10.1073/pnas.84.15.5192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sikorska M., Whitfield J. F., Walker P. R. The regulatory and catalytic subunits of cAMP-dependent protein kinases are associated with transcriptionally active chromatin during changes in gene expression. J Biol Chem. 1988 Feb 25;263(6):3005–3011. [PubMed] [Google Scholar]
  23. Supattapone S., Danoff S. K., Theibert A., Joseph S. K., Steiner J., Snyder S. H. Cyclic AMP-dependent phosphorylation of a brain inositol trisphosphate receptor decreases its release of calcium. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8747–8750. doi: 10.1073/pnas.85.22.8747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tagliaferri P., Katsaros D., Clair T., Ally S., Tortora G., Neckers L., Rubalcava B., Parandoosh Z., Chang Y. A., Revankar G. R. Synergistic inhibition of growth of breast and colon human cancer cell lines by site-selective cyclic AMP analogues. Cancer Res. 1988 Mar 15;48(6):1642–1650. [PubMed] [Google Scholar]
  25. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Walsh D. A., Perkins J. P., Krebs E. G. An adenosine 3',5'-monophosphate-dependant protein kinase from rabbit skeletal muscle. J Biol Chem. 1968 Jul 10;243(13):3763–3765. [PubMed] [Google Scholar]
  27. Wu J. C., Wang J. H. Sequence-selective DNA binding to the regulatory subunit of cAMP-dependent protein kinase. J Biol Chem. 1989 Jun 15;264(17):9989–9993. [PubMed] [Google Scholar]
  28. Yasui W., Sumiyoshi H., Ochiai A., Yamahara M., Tahara E. Type I and II cyclic adenosine 3':5'-monophosphate-dependent protein kinase in human gastric mucosa and carcinomas. Cancer Res. 1985 Apr;45(4):1565–1568. [PubMed] [Google Scholar]

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