Abstract
Using a small cell lung cancer (SCLC) cDNA library, we obtained clones for the creatine kinase-B (CK-B) gene and determined the nucleotide sequence for the protein coding and 3' untranslated region (3' UT). The human translated protein spans 381 residues and the amino acid homology with rabbit CK-B is greater than 98%. We have demonstrated that a nucleic acid probe encompassing the protein coding region will also hybridize to CK-M sequences while a probe derived from the 3' UT region is CK-B specific. When a B-isoenzyme specific sequence is hybridized to Eco RI cut genomic DNA, two independent restriction fragment polymorphisms are detected. We have subsequently localized these two CK-B homologous sequences to chromosomes 14q32 and 16. Finally, we show that increased levels of CK-B seen in SCLC are not accompanied by gene amplification or rearrangement, but reflect a greatly enhanced level of CK-B specific mRNA that is not seen in non-SCLC lines thus far examined.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benfield P. A., Zivin R. A., Miller L. S., Sowder R., Smythers G. W., Henderson L., Oroszlan S., Pearson M. L. Isolation and sequence analysis of cDNA clones coding for rat skeletal muscle creatine kinase. J Biol Chem. 1984 Dec 10;259(23):14979–14984. [PubMed] [Google Scholar]
- Bessman S. P., Carpenter C. L. The creatine-creatine phosphate energy shuttle. Annu Rev Biochem. 1985;54:831–862. doi: 10.1146/annurev.bi.54.070185.004151. [DOI] [PubMed] [Google Scholar]
- COHEN S. Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the new-born animal. J Biol Chem. 1962 May;237:1555–1562. [PubMed] [Google Scholar]
- Cande W. Z. Creatine kinase role in anaphase chromosome movement. Nature. 1983 Aug 11;304(5926):557–558. doi: 10.1038/304557a0. [DOI] [PubMed] [Google Scholar]
- Carney D. N., Gazdar A. F., Bepler G., Guccion J. G., Marangos P. J., Moody T. W., Zweig M. H., Minna J. D. Establishment and identification of small cell lung cancer cell lines having classic and variant features. Cancer Res. 1985 Jun;45(6):2913–2923. [PubMed] [Google Scholar]
- Carpenter C. L., Mohan C., Bessman S. P. Inhibition of protein and lipid synthesis in muscle by 2,4-dinitrofluorobenzene, an inhibitor of creatine phosphokinase. Biochem Biophys Res Commun. 1983 Mar 29;111(3):884–889. doi: 10.1016/0006-291x(83)91382-7. [DOI] [PubMed] [Google Scholar]
- Chern C. J., Tan P., Park H. Chromosomal mapping of human creatine kinase (brain type) using human-rodent somatic cell hybrids. Cytogenet Cell Genet. 1980;27(4):232–237. doi: 10.1159/000131491. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Cooper T. A., Ordahl C. P. A single cardiac troponin T gene generates embryonic and adult isoforms via developmentally regulated alternate splicing. J Biol Chem. 1985 Sep 15;260(20):11140–11148. [PubMed] [Google Scholar]
- Eppenberger M. E., Eppenberger H. M., Kaplan N. O. Evolution of creatine kinase. Nature. 1967 Apr 15;214(5085):239–241. doi: 10.1038/214239a0. [DOI] [PubMed] [Google Scholar]
- Gazdar A. F., Zweig M. H., Carney D. N., Van Steirteghen A. C., Baylin S. B., Minna J. D. Levels of creatine kinase and its BB isoenzyme in lung cancer specimens and cultures. Cancer Res. 1981 Jul;41(7):2773–2777. [PubMed] [Google Scholar]
- Hossle J. P., Rosenberg U. B., Schäfer B., Eppenberger H. M., Perriard J. C. The primary structure of chicken B-creatine kinase and evidence for heterogeneity of its mRNA. Nucleic Acids Res. 1986 Feb 11;14(3):1449–1463. doi: 10.1093/nar/14.3.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lebherz H. G., Burke T., Shackelford J. E., Strickler J. E., Wilson K. J. Specific proteolytic modification of creatine kinase isoenzymes. Implication of C-terminal involvement in enzymic activity but not in subunit-subunit recognition. Biochem J. 1986 Jan 1;233(1):51–56. doi: 10.1042/bj2330051. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
- Levitsky D. O., Levchenko T. S., Saks V. A., Sharov V. G., Smirnov V. N. The role of creatine phosphokinase in supplying energy for the calcium pump system of heart sarcoplasmic reticulum. Membr Biochem. 1978;2(1):81–96. doi: 10.3109/09687687809063859. [DOI] [PubMed] [Google Scholar]
- Loike J. D., Kozler V. F., Silverstein S. C. Increased ATP and creatine phosphate turnover in phagocytosing mouse peritoneal macrophages. J Biol Chem. 1979 Oct 10;254(19):9558–9564. [PubMed] [Google Scholar]
- McBride O. W., Battey J., Hollis G. F., Swan D. C., Siebenlist U., Leder P. Localization of human variable and constant region immunoglobulin heavy chain genes on subtelomeric band q32 of chromosome 14. Nucleic Acids Res. 1982 Dec 20;10(24):8155–8170. doi: 10.1093/nar/10.24.8155. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McBride O. W., Hieter P. A., Hollis G. F., Swan D., Otey M. C., Leder P. Chromosomal location of human kappa and lambda immunoglobulin light chain constant region genes. J Exp Med. 1982 May 1;155(5):1480–1490. doi: 10.1084/jem.155.5.1480. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
- Myers R. M., Larin Z., Maniatis T. Detection of single base substitutions by ribonuclease cleavage at mismatches in RNA:DNA duplexes. Science. 1985 Dec 13;230(4731):1242–1246. doi: 10.1126/science.4071043. [DOI] [PubMed] [Google Scholar]
- Nonessentiality of the active sulfhydryl group of rabbit muscle creatine kinase. J Biol Chem. 1974 May 25;249(10):3317–3318. [PubMed] [Google Scholar]
- Ordahl C. P., Evans G. L., Cooper T. A., Kunz G., Perriard J. C. Complete cDNA-derived amino acid sequence of chick muscle creatine kinase. J Biol Chem. 1984 Dec 25;259(24):15224–15227. [PubMed] [Google Scholar]
- Ottaway J. H. Evidence for binding of cytoplasmic creatine kinase to structural elements in heart muscle. Nature. 1967 Jul 29;215(5100):521–522. doi: 10.1038/215521b0. [DOI] [PubMed] [Google Scholar]
- Pickering L., Pang H., Biemann K., Munro H., Schimmel P. Two tissue-specific isozymes of creatine kinase have closely matched amino acid sequences. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2310–2314. doi: 10.1073/pnas.82.8.2310. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Polsky F., Edgell M. H., Seidman J. G., Leder P. High capacity gel preparative electrophoresis for purification of fragments of genomic DNA. Anal Biochem. 1978 Jul 1;87(2):397–410. doi: 10.1016/0003-2697(78)90689-9. [DOI] [PubMed] [Google Scholar]
- Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
- Roman D., Billadello J., Gordon J., Grace A., Sobel B., Strauss A. Complete nucleotide sequence of dog heart creatine kinase mRNA: conservation of amino acid sequence within and among species. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8394–8398. doi: 10.1073/pnas.82.24.8394. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg U. B., Eppenberger H. M., Perriard J. C. Occurrence of heterogenous forms of the subunits of creatine kinase in various muscle and nonmuscle tissues and their behaviour during myogenesis. Eur J Biochem. 1981 May;116(1):87–92. doi: 10.1111/j.1432-1033.1981.tb05304.x. [DOI] [PubMed] [Google Scholar]
- Rosenberg U. B., Kunz G., Frischauf A., Lehrach H., Mähr R., Eppenberger H. M., Perriard J. C. Molecular cloning and expression during myogenesis of sequences coding for M-creatine kinase. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6589–6592. doi: 10.1073/pnas.79.21.6589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sausville E. A., Lebacq-Verheyden A. M., Spindel E. R., Cuttitta F., Gazdar A. F., Battey J. F. Expression of the gastrin-releasing peptide gene in human small cell lung cancer. Evidence for alternative processing resulting in three distinct mRNAs. J Biol Chem. 1986 Feb 15;261(5):2451–2457. [PubMed] [Google Scholar]
- Schmidt C. J., Hamer D. H., McBride O. W. Chromosomal location of human metallothionein genes: implications for Menkes' disease. Science. 1984 Jun 8;224(4653):1104–1106. doi: 10.1126/science.6719135. [DOI] [PubMed] [Google Scholar]
- Schweinfest C. W., Kwiatkowski R. W., Dottin R. P. Molecular cloning of a DNA sequence complementary to creatine kinase M mRNA from chickens. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4997–5000. doi: 10.1073/pnas.79.16.4997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Tombes R. M., Shapiro B. M. Metabolite channeling: a phosphorylcreatine shuttle to mediate high energy phosphate transport between sperm mitochondrion and tail. Cell. 1985 May;41(1):325–334. doi: 10.1016/0092-8674(85)90085-6. [DOI] [PubMed] [Google Scholar]
- Watts D. C., Focant B., Moreland B. M., Watts R. L. Formation of a hybrid enzyme between echinoderm arginine kinase and mammalian creatine kinase. Nat New Biol. 1972 May 10;237(71):51–53. doi: 10.1038/newbio237051a0. [DOI] [PubMed] [Google Scholar]
- Weil D., Van Cong N., Gross M. S., Foubert C., Frézal J. Localisation du gène de la créatine kinase, BB sur le chromosome 14 par l'analyse des hybrides homme-rongeur. Ann Genet. 1980;23(3):150–154. [PubMed] [Google Scholar]
- West B. L., Babbitt P. C., Mendez B., Baxter J. D. Creatine kinase protein sequence encoded by a cDNA made from Torpedo californica electric organ mRNA. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7007–7011. doi: 10.1073/pnas.81.22.7007. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winter E., Yamamoto F., Almoguera C., Perucho M. A method to detect and characterize point mutations in transcribed genes: amplification and overexpression of the mutant c-Ki-ras allele in human tumor cells. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7575–7579. doi: 10.1073/pnas.82.22.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]