Abstract
For the isolation of cDNA clones encoding the carcinoembryonic antigen (CEA), we have constructed a cDNA library from human colon tumor mRNA. The library was screened with various oligonucleotides whose sequence had been deduced from partial amino acid sequence data for CEA. Positive candidate clones were hybridized with a probe for repetitive DNA, because CEA mRNA contains an Alu repetitive element, and with a fragment of a genomic clone of nonspecific cross-reacting antigen, an antigen closely related to CEA. Here we report the nucleotide sequence of the two overlapping CEA cDNA clones comprising 1422 nucleotides of CEA mRNA. This sequence encodes the 372 COOH-terminal amino acids of CEA followed by 305 nucleotides of 3' untranslated sequence containing a truncated Alu repeat. The predicted protein sequence is composed of two repeats comprising 178 amino acids, each with an exceptionally high homology of 67%. Each repeat unit contains four conserved cysteine residues and six to nine putative N-glycosylation sites. CEA mRNA is most strongly expressed in primary colon tumors and, to a lesser extent, in normal colonic tissue. No CEA mRNA is found in HeLa cells and normal human fibroblasts.
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.
- Abuharfeil N., Grunert F., von Kleist S. Non-specific crossreacting antigen (NCA) does not contain methionine. Tumour Biol. 1984;5(6):339–350. [PubMed] [Google Scholar]
- Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Buchegger F., Schreyer M., Carrel S., Mach J. P. Monoclonal antibodies identify a CEA crossreacting antigen of 95 kD (NCA-95) distinct in antigenicity and tissue distribution from the previously described NCA of 55 kD. Int J Cancer. 1984 May 15;33(5):643–649. doi: 10.1002/ijc.2910330515. [DOI] [PubMed] [Google Scholar]
- Chandrasekaran E. V., Davila M., Nixon D. W., Goldfarb M., Mendicino J. Isolation and structures of the oligosaccharide units of carcinoembryonic antigen. J Biol Chem. 1983 Jun 10;258(11):7213–7222. [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Feng D. F., Johnson M. S., Doolittle R. F. Aligning amino acid sequences: comparison of commonly used methods. J Mol Evol. 1984;21(2):112–125. doi: 10.1007/BF02100085. [DOI] [PubMed] [Google Scholar]
- Fiddes J. C., Goodman H. M. Isolation, cloning and sequence analysis of the cDNA for the alpha-subunit of human chorionic gonadotropin. Nature. 1979 Oct 4;281(5730):351–356. doi: 10.1038/281351a0. [DOI] [PubMed] [Google Scholar]
- Fritsche R., Mach J. P. Isolation and characterization of carcinoembryonic antigen (CEA) extracted from normal human colon mucosa. Immunochemistry. 1977 Feb;14(2):119–127. doi: 10.1016/0019-2791(77)90290-7. [DOI] [PubMed] [Google Scholar]
- Gilbert W. Why genes in pieces? Nature. 1978 Feb 9;271(5645):501–501. doi: 10.1038/271501a0. [DOI] [PubMed] [Google Scholar]
- Gold P., Krupey J., Ansari H. Position of the carcinoembryonic antigen of the human digestive system in ultrastructure of tumor cell surface. J Natl Cancer Inst. 1970 Aug;45(2):219–225. [PubMed] [Google Scholar]
- Gold P., Shuster J. Historical development and potential uses of tumor antigens as markers of human cancer growth. Cancer Res. 1980 Aug;40(8 Pt 2):2973–2976. [PubMed] [Google Scholar]
- Goldenberg D. M., DeLand F., Kim E., Bennett S., Primus F. J., van Nagell J. R., Jr, Estes N., DeSimone P., Rayburn P. Use of radiolabeled antibodies to carcinoembryonic antigen for the detection and localization of diverse cancers by external photoscanning. N Engl J Med. 1978 Jun 22;298(25):1384–1386. doi: 10.1056/NEJM197806222982503. [DOI] [PubMed] [Google Scholar]
- Goldenberg D. M., Gaffar S. A., Bennett S. J., Beach J. L. Experimental radioimmunotherapy of a xenografted human colonic tumor (GW-39) producing carcinoembryonic antigen. Cancer Res. 1981 Nov;41(11 Pt 1):4354–4360. [PubMed] [Google Scholar]
- Grunert F., AbuHarfeil N., Schwarz K., von Kleist S. Two CEA and three NCA species, although distinguishable by monoclonal antibodies, have nearly identical peptide patterns. Int J Cancer. 1985 Sep 15;36(3):357–362. [PubMed] [Google Scholar]
- Grunert F., Luckenbach G. A., Haderlie B., Schwarz K., von Kleist S. Comparison of colon-, lung-, and breast-derived carcinoembryonic antigen and cross-reacting antigens by monoclonal antibodies and fingerprint analysis. Ann N Y Acad Sci. 1983;417:75–85. doi: 10.1111/j.1749-6632.1983.tb32851.x. [DOI] [PubMed] [Google Scholar]
- Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
- Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
- Hedin A., Hammarström S., Larsson A. Specificities and binding properties of eight monoclonal antibodies against carcinoembryonic antigen. Mol Immunol. 1982 Dec;19(12):1641–1648. doi: 10.1016/0161-5890(82)90275-9. [DOI] [PubMed] [Google Scholar]
- Hood L., Steinmetz M., Malissen B. Genes of the major histocompatibility complex of the mouse. Annu Rev Immunol. 1983;1:529–568. doi: 10.1146/annurev.iy.01.040183.002525. [DOI] [PubMed] [Google Scholar]
- Ishioka N., Takahashi N., Putnam F. W. Amino acid sequence of human plasma alpha 1B-glycoprotein: homology to the immunoglobulin supergene family. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2363–2367. doi: 10.1073/pnas.83.8.2363. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jelinek W. R., Schmid C. W. Repetitive sequences in eukaryotic DNA and their expression. Annu Rev Biochem. 1982;51:813–844. doi: 10.1146/annurev.bi.51.070182.004121. [DOI] [PubMed] [Google Scholar]
- Jothy S., Brazinsky S. A., Chin-A-Loy M., Haggarty A., Krantz M. J., Cheung M., Fuks A. Characterization of monoclonal antibodies to carcinoembryonic antigen with increased tumor specificity. Lab Invest. 1986 Jan;54(1):108–117. [PubMed] [Google Scholar]
- Kessler M. J., Shively J. E., Pritchard D. G., Todd C. W. Isolation, immunological characterization, and structural studies of a tumor antigen related to carcinoembryonic antigen. Cancer Res. 1978 Apr;38(4):1041–1048. [PubMed] [Google Scholar]
- Kornblihtt A. R., Umezawa K., Vibe-Pedersen K., Baralle F. E. Primary structure of human fibronectin: differential splicing may generate at least 10 polypeptides from a single gene. EMBO J. 1985 Jul;4(7):1755–1759. doi: 10.1002/j.1460-2075.1985.tb03847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuroki M., Shinoda T., Takayasu T., Koga Y., Matsuoka Y. Immunological characterization and structural studies of normal fecal antigen-1 related to carcinoembryonic antigen. Mol Immunol. 1982 Mar;19(3):399–406. doi: 10.1016/0161-5890(82)90205-x. [DOI] [PubMed] [Google Scholar]
- Lathe R. Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations. J Mol Biol. 1985 May 5;183(1):1–12. doi: 10.1016/0022-2836(85)90276-1. [DOI] [PubMed] [Google Scholar]
- Matlashewski G., Lamb P., Pim D., Peacock J., Crawford L., Benchimol S. Isolation and characterization of a human p53 cDNA clone: expression of the human p53 gene. EMBO J. 1984 Dec 20;3(13):3257–3262. doi: 10.1002/j.1460-2075.1984.tb02287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
- Pagé M., Delorme F., Lafontaine F., Dumas L. Chemotherapy with daunorubicin-anti-CEA conjugates in human colon adenocarcinoma grafted in nude mice. Semin Oncol. 1984 Dec;11(4 Suppl 3):56–58. [PubMed] [Google Scholar]
- Paxton R. J., Mooser G., Pande H., Lee T. D., Shively J. E. Sequence analysis of carcinoembryonic antigen: identification of glycosylation sites and homology with the immunoglobulin supergene family. Proc Natl Acad Sci U S A. 1987 Feb;84(4):920–924. doi: 10.1073/pnas.84.4.920. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rüther U., Koenen M., Otto K., Müller-Hill B. pUR222, a vector for cloning and rapid chemical sequencing of DNA. Nucleic Acids Res. 1981 Aug 25;9(16):4087–4098. doi: 10.1093/nar/9.16.4087. [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]
- Shi Z. R., Tsao D., Kim Y. S. Subcellular distribution, synthesis, and release of carcinoembryonic antigen in cultured human colon adenocarcinoma cell lines. Cancer Res. 1983 Sep;43(9):4045–4049. [PubMed] [Google Scholar]
- Shively J. E., Todd C. W., Go V. L., Egan M. L. Amino-terminal sequence of a carcinoembryonic antigen-like glycoprotein isolated from the colonic lavages of healthy individuals. Cancer Res. 1978 Mar;38(3):503–505. [PubMed] [Google Scholar]
- Terry W. D., Henkart P. A., Coligan J. E., Todd C. W. Carcinoembryonic antigen: characterization and clinical applications. Transplant Rev. 1974;20(0):100–129. doi: 10.1111/j.1600-065x.1974.tb00143.x. [DOI] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson J. A., Pande H., Paxton R. J., Shively L., Padma A., Simmer R. L., Todd C. W., Riggs A. D., Shively J. E. Molecular cloning of a gene belonging to the carcinoembryonic antigen gene family and discussion of a domain model. Proc Natl Acad Sci U S A. 1987 May;84(9):2965–2969. doi: 10.1073/pnas.84.9.2965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Whitehead A. S., Goldberger G., Woods D. E., Markham A. F., Colten H. R. Use of a cDNA clone for the fourth component of human complement (C4) for analysis of a genetic deficiency of C4 in guinea pig. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5387–5391. doi: 10.1073/pnas.80.17.5387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu B., Hunt C., Morimoto R. Structure and expression of the human gene encoding major heat shock protein HSP70. Mol Cell Biol. 1985 Feb;5(2):330–341. doi: 10.1128/mcb.5.2.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yamamoto T., Davis C. G., Brown M. S., Schneider W. J., Casey M. L., Goldstein J. L., Russell D. W. The human LDL receptor: a cysteine-rich protein with multiple Alu sequences in its mRNA. Cell. 1984 Nov;39(1):27–38. doi: 10.1016/0092-8674(84)90188-0. [DOI] [PubMed] [Google Scholar]
- Zimmermann W., Friedrich R., Grunert F., Luckenbach G. A., Thompson J., von Kleist S. Characterization of messenger RNA specific for carcinoembryonic antigen. Ann N Y Acad Sci. 1983;417:21–30. doi: 10.1111/j.1749-6632.1983.tb32844.x. [DOI] [PubMed] [Google Scholar]