Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Jul;84(13):4552–4556. doi: 10.1073/pnas.84.13.4552

Cloning of a brain protein identified by autoantibodies from a patient with paraneoplastic cerebellar degeneration.

E J Dropcho, Y T Chen, J B Posner, L J Old
PMCID: PMC305128  PMID: 3474619

Abstract

Autoantibodies directed against neuronal proteins have been identified in some patients with paraneoplastic cerebellar degeneration. To identify the molecular targets for these autoantibodies, we constructed a lambda gt11 cDNA expression library from human cerebellum and screened the library with IgG from a patient with paraneoplastic cerebellar degeneration. A single clone, pCDR2, produced a fusion protein that reacted strongly with the patient's IgG. The isolated pCDR2 clone was used to identify six overlapping cDNA clones. Sequencing of the pCDR clones revealed a distinctive pattern consisting of a unit of 18 nucleotides (6 amino acids) repeated in tandem along the entire cDNA sequence. This sequence is unlike any previously described eukaryotic gene. Southern blot analysis was consistent with single-copy representation of the CDR (cerebellar degeneration-related) gene in the human and mouse genome. RNA blotting studies with normal tissues showed expression of the CDR gene to be largely restricted to brain. Expression of the CDR message was also noted in cell lines derived from cancers of neuroectodermal, kidney, and lung origin.

Full text

PDF
4552

Images in this article

4553Image
on p.4553
4553Image
on p.4553
4555Image
on p.4555
4555Image
on p.4555
4555Image
on p.4555
4555Image
on p.4555
4555Image
on p.4555
4555Image
on p.4555

Selected References

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

  1. Allison L. A., Moyle M., Shales M., Ingles C. J. Extensive homology among the largest subunits of eukaryotic and prokaryotic RNA polymerases. Cell. 1985 Sep;42(2):599–610. doi: 10.1016/0092-8674(85)90117-5. [DOI] [PubMed] [Google Scholar]
  2. Brain L., Wilkinson M. Subacute cerebellar degeneration associated with neoplasms. Brain. 1965 Sep;88(3):465–478. doi: 10.1093/brain/88.3.465. [DOI] [PubMed] [Google Scholar]
  3. Chen Y. T., Obata Y., Stockert E., Old L. J. Thymus-leukemia (TL) antigens of the mouse. Analysis of TL mRNA and TL cDNA TL+ and TL- strains. J Exp Med. 1985 Oct 1;162(4):1134–1148. doi: 10.1084/jem.162.4.1134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  5. Corden J. L., Cadena D. L., Ahearn J. M., Jr, Dahmus M. E. A unique structure at the carboxyl terminus of the largest subunit of eukaryotic RNA polymerase II. Proc Natl Acad Sci U S A. 1985 Dec;82(23):7934–7938. doi: 10.1073/pnas.82.23.7934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cunningham J., Graus F., Anderson N., Posner J. B. Partial characterization of the Purkinje cell antigens in paraneoplastic cerebellar degeneration. Neurology. 1986 Sep;36(9):1163–1168. doi: 10.1212/wnl.36.9.1163. [DOI] [PubMed] [Google Scholar]
  7. Dame J. B., Williams J. L., McCutchan T. F., Weber J. L., Wirtz R. A., Hockmeyer W. T., Maloy W. L., Haynes J. D., Schneider I., Roberts D. Structure of the gene encoding the immunodominant surface antigen on the sporozoite of the human malaria parasite Plasmodium falciparum. Science. 1984 Aug 10;225(4662):593–599. doi: 10.1126/science.6204383. [DOI] [PubMed] [Google Scholar]
  8. DeLozanne A., Lewis M., Spudich J. A., Leinwand L. A. Cloning and characterization of a nonmuscle myosin heavy chain cDNA. Proc Natl Acad Sci U S A. 1985 Oct;82(20):6807–6810. doi: 10.1073/pnas.82.20.6807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eckert R. L., Green H. Structure and evolution of the human involucrin gene. Cell. 1986 Aug 15;46(4):583–589. doi: 10.1016/0092-8674(86)90884-6. [DOI] [PubMed] [Google Scholar]
  10. Ellis J., Ozaki L. S., Gwadz R. W., Cochrane A. H., Nussenzweig V., Nussenzweig R. S., Godson G. N. Cloning and expression in E. coli of the malarial sporozoite surface antigen gene from Plasmodium knowlesi. Nature. 1983 Apr 7;302(5908):536–538. doi: 10.1038/302536a0. [DOI] [PubMed] [Google Scholar]
  11. Fisher D. Z., Chaudhary N., Blobel G. cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6450–6454. doi: 10.1073/pnas.83.17.6450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gallin W. J., Prediger E. A., Edelman G. M., Cunningham B. A. Isolation of a cDNA clone for the liver cell adhesion molecule (L-CAM). Proc Natl Acad Sci U S A. 1985 May;82(9):2809–2813. doi: 10.1073/pnas.82.9.2809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Greenlee J. E., Brashear H. R. Antibodies to cerebellar Purkinje cells in patients with paraneoplastic cerebellar degeneration and ovarian carcinoma. Ann Neurol. 1983 Dec;14(6):609–613. doi: 10.1002/ana.410140603. [DOI] [PubMed] [Google Scholar]
  14. Greenlee J. E., Lipton H. L. Anticerebellar antibodies in serum and cerebrospinal fluid of a patient with oat cell carcinoma of the lung and paraneoplastic cerebellar degeneration. Ann Neurol. 1986 Jan;19(1):82–85. doi: 10.1002/ana.410190117. [DOI] [PubMed] [Google Scholar]
  15. Jaeckle K. A., Graus F., Houghton A., Cardon-Cardo C., Nielsen S. L., Posner J. B. Autoimmune response of patients with paraneoplastic cerebellar degeneration to a Purkinje cell cytoplasmic protein antigen. Ann Neurol. 1985 Nov;18(5):592–600. doi: 10.1002/ana.410180513. [DOI] [PubMed] [Google Scholar]
  16. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Locht C., Chesebro B., Race R., Keith J. M. Molecular cloning and complete sequence of prion protein cDNA from mouse brain infected with the scrapie agent. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6372–6376. doi: 10.1073/pnas.83.17.6372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marchuk D., McCrohon S., Fuchs E. Remarkable conservation of structure among intermediate filament genes. Cell. 1984 Dec;39(3 Pt 2):491–498. doi: 10.1016/0092-8674(84)90456-2. [DOI] [PubMed] [Google Scholar]
  20. Obata Y., Chen Y. T., Stockert E., Old L. J. Structural analysis of TL genes of the mouse. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5475–5479. doi: 10.1073/pnas.82.16.5475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Oesch B., Westaway D., Wälchli M., McKinley M. P., Kent S. B., Aebersold R., Barry R. A., Tempst P., Teplow D. B., Hood L. E. A cellular gene encodes scrapie PrP 27-30 protein. Cell. 1985 Apr;40(4):735–746. doi: 10.1016/0092-8674(85)90333-2. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Stone D., Smillie L. B. The amino acid sequence of rabbit skeletal alpha-tropomyosin. The NH2-terminal half and complete sequence. J Biol Chem. 1978 Feb 25;253(4):1137–1148. [PubMed] [Google Scholar]
  25. Tanaka K., Yamazaki M., Sato S., Toyoshima I., Yamamoto A., Miyatake T. Antibodies to brain proteins in paraneoplastic cerebellar degeneration. Neurology. 1986 Sep;36(9):1169–1172. doi: 10.1212/wnl.36.9.1169. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Trotter J. L., Hendin B. A., Osterland C. K. Cerebellar degeneration with Hodgkin disease. An immunological study. Arch Neurol. 1976 Sep;33(9):660–661. doi: 10.1001/archneur.1976.00500090066014. [DOI] [PubMed] [Google Scholar]
  28. White B. A., Bancroft F. C. Cytoplasmic dot hybridization. Simple analysis of relative mRNA levels in multiple small cell or tissue samples. J Biol Chem. 1982 Aug 10;257(15):8569–8572. [PubMed] [Google Scholar]
  29. Young R. A., Davis R. W. Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1194–1198. doi: 10.1073/pnas.80.5.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES