1887

Abstract

By screening a cDNA library with a cDNA encoding the cAMP-binding protein CABP1, under conditions of reduced stringency, we have isolated clones which code for two closely related molecules. Hybrid selection experiments indicated that these cDNAs encoded polypeptides with molecular masses of 34 (p34) and 31 (p31) kDa, both of which were recognized by anti-CABP1 monoclonal antibodies. Sequence analysis revealed that the clones were identical except for the presence of a 102 nucleotide segment inserted in-frame in the p34 cDNAs, just downstream of the translation initiation codon. DNA blot analysis suggested that p34 and p31 were encoded by the same gene. This hypothesis was strongly supported by the observation that both polypeptides were generated when a single cDNA was expressed under the control of the actin 15 promoter in cells. RNA blot analysis indicated that the cDNAs were complementary to three developmentally regulated transcripts of sizes 1.15 kb, 1.25 kb and 1.4 kb. Comparison of the derived amino acid sequences of p34 and p31 with those of the two subunits of CABP1 indicated that these polypeptides were very closely related, and that the corresponding genes probably arose by duplication followed by sequence divergence. Finally, the carboxy termini of these four polypeptides demonstrated 50% similarity to two polypeptides encoded by a bacterial plasmid which confers resistance to tellurium anions.

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/content/journal/micro/10.1099/00221287-137-3-501
1991-03-01
2021-08-04
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References

  1. Early A. E., Williams J. G. 1987; Two vectors which facilitate gene manipulation and a simplified transformation procedure for Dictyostelium discoideum. Gene 59:99–106
    [Google Scholar]
  2. Firtel R. A., Bonner J. 1972; Characterization of the genome of the cellular slime mold Dictyostelium discoideum. Journal of Molecular Biology 66:339–361
    [Google Scholar]
  3. Firtel R. A., Van Haastert P. J. M., Kimmel A. R., Devreotes P. N. 1989; G protein linked signal transduction pathways in development: Dictyostelium as a model system. Cell 58:235–239
    [Google Scholar]
  4. Gerisch G. 1987; Cyclic AMP and other signals controlling cell development and differentiation in Dictyostelium. Annual Review of Biochemistry 56:553–579
    [Google Scholar]
  5. Grant C. E., Tsang A. 1990; Cloning and characterization of cDNAs encoding a novel cyclic AMP-binding protein in Dictyostelium discoideum. Gene (in the Press)
    [Google Scholar]
  6. Grant C. E., Bain G., Tsang A. 1990; The molecular basis for the alternative splicing of the CABP1 transcripts in Dictyostelium discoideum. Nucleic Acids Research 18:5457–5463
    [Google Scholar]
  7. Jobling M. G., Ritchie D. A. 1988; The nucleotide sequence of a plasmid determinant for resistance to tellurium anions. Gene 66:245–258
    [Google Scholar]
  8. Kay C. A., Noce T., Tsang A. S. 1987; Translocation of an unusual cAMP receptor to the nucleus during development of Dictyostelium discoideum. Proceedings of the National Academy of Sciences of the United States of America 842322–2326
    [Google Scholar]
  9. Kay R. R. 1989; Evidence that elevated intracellular cyclic AMP triggers spore maturation in Dictyostelium. Development 105:753–759
    [Google Scholar]
  10. Kimmel A. R. 1987; Different molecular mechanisms for cAMP regulation of gene expression during Dictyostelium development. Developmental Biology 122:163–171
    [Google Scholar]
  11. Klein P. S., Sun T. J., Saxe C. L., Kimmel A. R., Johnson R. L., Devreotes P. N. 1988; A chemoattractant receptor controls development in Dictyostelium discoideum. Science 241:1467–1472
    [Google Scholar]
  12. Kunkel T. A. 1985; Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences of the United States of America 82488–492
    [Google Scholar]
  13. Loomis W. F. 1982 The development of Dictyostelium discoideum New York: Academic Press;
    [Google Scholar]
  14. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  15. Pearson W. R., Lipman D. J. 1988; Improved tools for biological sequence analysis. Proceedings of the National Academy of Sciences of the United States of America 852444–2448
    [Google Scholar]
  16. Riley B. B., Jensen B. R., Barclay S. L. 1989; Conditions that elevate intracellular cAMP levels promote spore formation in Dictyostelium. Differentiation 41:5–13
    [Google Scholar]
  17. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 745463–5467
    [Google Scholar]
  18. Tsang A. S., Grant C., Kay C., Bain G., Greenwood M., Noce T., Tasaka M. 1988; Characterization of an unusual cAMP receptor and its related polypeptides in Dictyostelium discoideum. Developmental Genetics 9:237–245
    [Google Scholar]
  19. Tsang A. S., Tasaka M. 1986; Identification of multiple cyclic AMP binding proteins in developing Dictyostelium discoideum cells. Journal of Biological Chemistry 261:10753–10759
    [Google Scholar]
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