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. 1989 Dec 25;17(24):10321–10335. doi: 10.1093/nar/17.24.10321

Primary structure and developmental expression of a large cytoplasmic domain form of Xenopus laevis neural cell adhesion molecule (NCAM).

P A Krieg 1, D S Sakaguchi 1, C R Kintner 1
PMCID: PMC335303  PMID: 2481269

Abstract

The neural cell adhesion molecule, (NCAM), is involved in cell-cell interactions during development of the vertebrate nervous system. NCAM exists in multiple protein forms and these are selectively expressed in different cells and at different times during development. Here we report the complete amino acid sequence of the large cytoplasmic form of Xenopus laevis NCAM, derived from a full-length cDNA clone. Using specific probes the expression of different NCAM transcripts during Xenopus embryogenesis has been examined. We find that transcripts encoding the large cytoplasmic domain form of NCAM exist in maternal RNA and that these are the only significant NCAM transcripts present until late gastrula when transcripts encoding the small cytoplasmic domain form of NCAM are first detected. No RNA encoding the small surface domain form of NCAM is detected during early development. These results indicate that the expression of NCAM sequences during early development of Xenopus differs from that described in other species.

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Selected References

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  1. Barbas J. A., Chaix J. C., Steinmetz M., Goridis C. Differential splicing and alternative polyadenylation generates distinct NCAM transcripts and proteins in the mouse. EMBO J. 1988 Mar;7(3):625–632. doi: 10.1002/j.1460-2075.1988.tb02856.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barthels D., Santoni M. J., Wille W., Ruppert C., Chaix J. C., Hirsch M. R., Fontecilla-Camps J. C., Goridis C. Isolation and nucleotide sequence of mouse NCAM cDNA that codes for a Mr 79,000 polypeptide without a membrane-spanning region. EMBO J. 1987 Apr;6(4):907–914. doi: 10.1002/j.1460-2075.1987.tb04837.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cole G. J., Loewy A., Cross N. V., Akeson R., Glaser L. Topographic localization of the heparin-binding domain of the neural cell adhesion molecule N-CAM. J Cell Biol. 1986 Nov;103(5):1739–1744. doi: 10.1083/jcb.103.5.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cunningham B. A., Hemperly J. J., Murray B. A., Prediger E. A., Brackenbury R., Edelman G. M. Neural cell adhesion molecule: structure, immunoglobulin-like domains, cell surface modulation, and alternative RNA splicing. Science. 1987 May 15;236(4803):799–806. doi: 10.1126/science.3576199. [DOI] [PubMed] [Google Scholar]
  5. Dickson G., Gower H. J., Barton C. H., Prentice H. M., Elsom V. L., Moore S. E., Cox R. D., Quinn C., Putt W., Walsh F. S. Human muscle neural cell adhesion molecule (N-CAM): identification of a muscle-specific sequence in the extracellular domain. Cell. 1987 Sep 25;50(7):1119–1130. doi: 10.1016/0092-8674(87)90178-4. [DOI] [PubMed] [Google Scholar]
  6. Edelman G. M. Morphoregulatory molecules. Biochemistry. 1988 May 17;27(10):3533–3543. doi: 10.1021/bi00410a001. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Frelinger A. L., 3rd, Rutishauser U. Topography of N-CAM structural and functional determinants. II. Placement of monoclonal antibody epitopes. J Cell Biol. 1986 Nov;103(5):1729–1737. doi: 10.1083/jcb.103.5.1729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goridis C., Deagostini-Bazin H., Hirn M., Hirsch M. R., Rougon G., Sadoul R., Langley O. K., Gombos G., Finne J. Neural surface antigens during nervous system development. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):527–537. doi: 10.1101/sqb.1983.048.01.057. [DOI] [PubMed] [Google Scholar]
  10. Goridis C., Hirn M., Santoni M. J., Gennarini G., Deagostini-Bazin H., Jordan B. R., Kiefer M., Steinmetz M. Isolation of mouse N-CAM-related cDNA: detection and cloning using monoclonal antibodies. EMBO J. 1985 Mar;4(3):631–635. doi: 10.1002/j.1460-2075.1985.tb03676.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hall A. K., Rutishauser U. Phylogeny of a neural cell adhesion molecule. Dev Biol. 1985 Jul;110(1):39–46. doi: 10.1016/0012-1606(85)90061-2. [DOI] [PubMed] [Google Scholar]
  12. He H. T., Barbet J., Chaix J. C., Goridis C. Phosphatidylinositol is involved in the membrane attachment of NCAM-120, the smallest component of the neural cell adhesion molecule. EMBO J. 1986 Oct;5(10):2489–2494. doi: 10.1002/j.1460-2075.1986.tb04526.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hemperly J. J., Edelman G. M., Cunningham B. A. cDNA clones of the neural cell adhesion molecule (N-CAM) lacking a membrane-spanning region consistent with evidence for membrane attachment via a phosphatidylinositol intermediate. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9822–9826. doi: 10.1073/pnas.83.24.9822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hemperly J. J., Murray B. A., Edelman G. M., Cunningham B. A. Sequence of a cDNA clone encoding the polysialic acid-rich and cytoplasmic domains of the neural cell adhesion molecule N-CAM. Proc Natl Acad Sci U S A. 1986 May;83(9):3037–3041. doi: 10.1073/pnas.83.9.3037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hoffman S., Sorkin B. C., White P. C., Brackenbury R., Mailhammer R., Rutishauser U., Cunningham B. A., Edelman G. M. Chemical characterization of a neural cell adhesion molecule purified from embryonic brain membranes. J Biol Chem. 1982 Jul 10;257(13):7720–7729. [PubMed] [Google Scholar]
  16. Jacobson M., Rutishauser U. Induction of neural cell adhesion molecule (NCAM) in Xenopus embryos. Dev Biol. 1986 Aug;116(2):524–531. doi: 10.1016/0012-1606(86)90153-3. [DOI] [PubMed] [Google Scholar]
  17. Kintner C. R., Melton D. A. Expression of Xenopus N-CAM RNA in ectoderm is an early response to neural induction. Development. 1987 Mar;99(3):311–325. doi: 10.1242/dev.99.3.311. [DOI] [PubMed] [Google Scholar]
  18. Kintner C. Effects of altered expression of the neural cell adhesion molecule, N-CAM, on early neural development in Xenopus embryos. Neuron. 1988 Sep;1(7):545–555. doi: 10.1016/0896-6273(88)90104-3. [DOI] [PubMed] [Google Scholar]
  19. Krieg P. A., Melton D. A. In vitro RNA synthesis with SP6 RNA polymerase. Methods Enzymol. 1987;155:397–415. doi: 10.1016/0076-6879(87)55027-3. [DOI] [PubMed] [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  21. Levi G., Crossin K. L., Edelman G. M. Expression sequences and distribution of two primary cell adhesion molecules during embryonic development of Xenopus laevis. J Cell Biol. 1987 Nov;105(5):2359–2372. doi: 10.1083/jcb.105.5.2359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Melton D. A., Cortese R. Transcription of cloned tRNA genes and the nuclear partitioning of a tRNA precursor. Cell. 1979 Dec;18(4):1165–1172. doi: 10.1016/0092-8674(79)90229-0. [DOI] [PubMed] [Google Scholar]
  23. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  24. Murray B. A., Hemperly J. J., Gallin W. J., MacGregor J. S., Edelman G. M., Cunningham B. A. Isolation of cDNA clones for the chicken neural cell adhesion molecule (N-CAM). Proc Natl Acad Sci U S A. 1984 Sep;81(17):5584–5588. doi: 10.1073/pnas.81.17.5584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murray B. A., Hemperly J. J., Prediger E. A., Edelman G. M., Cunningham B. A. Alternatively spliced mRNAs code for different polypeptide chains of the chicken neural cell adhesion molecule (N-CAM). J Cell Biol. 1986 Jan;102(1):189–193. doi: 10.1083/jcb.102.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nguyen C., Mattei M. G., Mattei J. F., Santoni M. J., Goridis C., Jordan B. R. Localization of the human NCAM gene to band q23 of chromosome 11: the third gene coding for a cell interaction molecule mapped to the distal portion of the long arm of chromosome 11. J Cell Biol. 1986 Mar;102(3):711–715. doi: 10.1083/jcb.102.3.711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Owens G. C., Edelman G. M., Cunningham B. A. Organization of the neural cell adhesion molecule (N-CAM) gene: alternative exon usage as the basis for different membrane-associated domains. Proc Natl Acad Sci U S A. 1987 Jan;84(1):294–298. doi: 10.1073/pnas.84.1.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sakaguchi D. S., Moeller J. F., Coffman C. R., Gallenson N., Harris W. A. Growth cone interactions with a glial cell line from embryonic Xenopus retina. Dev Biol. 1989 Jul;134(1):158–174. doi: 10.1016/0012-1606(89)90086-9. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Santoni M. J., Barthels D., Barbas J. A., Hirsch M. R., Steinmetz M., Goridis C., Wille W. Analysis of cDNA clones that code for the transmembrane forms of the mouse neural cell adhesion molecule (NCAM) and are generated by alternative RNA splicing. Nucleic Acids Res. 1987 Nov 11;15(21):8621–8641. doi: 10.1093/nar/15.21.8621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sunshine J., Balak K., Rutishauser U., Jacobson M. Changes in neural cell adhesion molecule (NCAM) structure during vertebrate neural development. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5986–5990. doi: 10.1073/pnas.84.16.5986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

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