Advertisement

Mammalian Genome

, Volume 5, Issue 11, pp 701–706 | Cite as

Cloning of a mouse protein kinase A catalytic subunit pseudogene and chromosomal mapping of C subunit isoforms

  • D. E. Cummings
  • S. Edelhoff
  • C. M. Disteche
  • G. S. McKnight
Original Contributions

Abstract

Two isoforms of the protein kinase A catalytic subunit, Cα and Cβ, have previously been described in the mouse. We now report the cloning and characterization of a novel C-related sequence, Cx, from a murine genomic library. Cx is 89.8% identical to part of the Cα coding region, but lacks all of the introns present in this gene, suggesting that is arose via retroposition. The existence of several frameshift mutations, premature termination codons, and missense mutations at critical sites confirms that it is a pseudogene. Furthermore, we are unable to detect any expression. Homology with functional protein kinase genes commences exactly at the first intron splice junction in Cα, downstream of the expected translational start codon. Cx is also truncated at its 3′ end by the interposition of two distinct, contiguous LINE-1 elements. By fluorescence in situ hybridization, we demonstrate that Cx is located on the X Chromosome (Chr), at band F3. This is displaced from its functional homologs, Cα and Cβ, which we map to mouse Chrs 8 (band C3) and 3 (band H3), respectively.

Keywords

Catalytic Subunit Frameshift Mutation Termination Codon Premature Termination Splice Junction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ashworth, A., Skene, B., Swift, S., Lovell-Badge, R. (1990). Zfa is an expressed retroposon derived from an alternative transcript of the Zfx gene. EMBO J. 9, 1529–1534.Google Scholar
  2. Battey, J., Max, E., McBride, W., Swan, D., Leder, P. (1982). A processed human immunoglobulin ɛ gene has moved to chromosome 9. Proc. Natl. Acad. Sci. 79, 5956–5960.Google Scholar
  3. Beebe, S., Oyen, O., Sandberg, M., Froysa, A., Hansson, V., Jahnsen, T. (1990) Molecular cloning of a tissue-specific protein kinase (Cγ) from human testis—representing a third isoform of the catalytic subunit of cAMP-dependent protein kinase. Mol. Endocrinol. 4, 465–475.Google Scholar
  4. Chrivia, J., Uhler, M., McKnight, G. (1988). Characterization of genomic clones coding for the Cα and Cβ subunits of mouse cAMP-dependent protein kinase. J. Biol. Chem. 263, 5739–5744.Google Scholar
  5. Dahl, H-H., Brown, R., Hutchison, W., Maragos, C., Brown, G. (1990). A testis-specific form of the human pyruvate dehydrogenase E1α subunit is coded for by an intronless gene on chromosome 4. Genomics 8, 225–232.Google Scholar
  6. Edelhoff, S., Ayer, D., Zervos, A., Steingrimess, E., Jenkins, N., Copeland, N., Eisenman, R., Brent, R., Disteche, C. (1994). Mapping of two genes encoding members of a distinct subfamily of MAX interacting proteins: MAD to human chromosome 2 and mouse chromosome 6, and MXI1 to human chromosome 10 and mouse chromosome 19. Oncogene 9, 665–668.Google Scholar
  7. Fourel, G., Trepo, C., Bougueleret, L., Henglein, B., Ponzetto, A., Tiollais, P., Buendia, M-A. (1990). Frequent activation of N-myc genes by hepadnavirus insertion in woodchuck liver tumours. Nature 347, 294–298.Google Scholar
  8. Green, M. (1989). Catalogue of mutant genes and polymorphic loci. In Genetic Variants and Strains of the Laboratory Mouse, M. Lyon, A. Searle, eds. (New York: Oxford University Press), pp. 12–403.Google Scholar
  9. Hanks, S., Quinn, A., Hunter, T. (1988). The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science 241, 42–52.Google Scholar
  10. Hollis, G., Hieter, P., McBride, O., Swan, D., Leder, P. (1982). Processed genes: a dispersed human immunoglobulin gene bearing evidence of RNA-type processing. Nature 296, 321–325.Google Scholar
  11. Jacq, C., Miller, J., Brownlee, G. (1977). A pseudogene structure in 5S DNA of Xenopus laevis. Cell 12, 109–120.Google Scholar
  12. Knighton, D., Zheng, J., Ten Eyck, L., Xuong, N., Taylor, S., Sowadski, J. (1991). Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. Science 253, 414–420.Google Scholar
  13. Kricker, M., Drake, J., Radman, M. (1992). Duplication-targeted DNA methylation and mutagenesis in the evolution of eukaryotic chromosomes. Proc. Natl. Acad. Sci. USA 89, 1075–1079.Google Scholar
  14. Maeda, N., Wu, C., Bliska, J., Reneke, J. (1988). Molecular evolution of intergenic DNA in higher primates: pattern of DNA changes, molecular clock and evolution of repetitive sequences. Mol. Biol. Evol. 5, 1–20.Google Scholar
  15. McCarrey, J., Thomas, K. (1987). Human testis-specific PGK gene lacks introns and possesses characteristics of a processed gene. Nature 326, 501–505.Google Scholar
  16. McKnight, G. (1991). Cyclic AMP second messenger systems. Curr. Opin. Cell Biol. 3, 213–217.Google Scholar
  17. Orellana, S., McKnight, G. (1990). The S49 kin- cell line transcribes and translates a functional mRNA coding for the catalytic subunit of cAMP-dependent protein kinase. J. Biol. Chem. 265, 3048–3053.Google Scholar
  18. Saiki, R., Gelfand, D., Stoffel, S., Scarf, S., Higuchi, R., Horn, G., Mullis, K., Erlich, H. (1988). Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487–491.Google Scholar
  19. Samuelson, L., Wiebauer, K., Snow, C., Meisler, M. (1990). Retroviral and pseudogene insertion sites reveal the lineage of human salivery and pancreatic amylase genes from a single gene during primate evolution. Mol. Cell. Biol. 10, 2513–2520.Google Scholar
  20. Scarpulla, R. (1985). Association of a truncated cytochrome c processed pseudogene with a similarly truncated member from a long interspersed repeat family of rat. Nucleic Acids Res. 13, 763–775.Google Scholar
  21. Shimada, T., Chen, M-J., Nienhuis, A. (1984). A human dihydrofolate reductase intronless pseudogene with an Alu repetitive sequence: multiple DNA insertions at a single chromosomal site. Gene 31, 1–8.Google Scholar
  22. Simard, J., Berube, D., Sandberg, M., Grzeschik, K-H., Gagne, R., Hansson, V., Jahnsen, T. (1992) Assignment of the gene encoding the catalytic subunit Cβ of cAMP-dependent protein kinase A to the p36 band on chromosome 1. Hum. Genet. 88, 653–657.Google Scholar
  23. Soares, M., Schon, E., Henderson, A., Karathanasis, S., Cate, R., Zeitlin, S., Chirgwin, J., Efstratiadis, A. (1985). RNA-mediated gene duplication: the rat preproinsulin I gene is a functional retroposon. Mol. Cell. Biol. 5, 2090–2103.Google Scholar
  24. Solberg, R., Sandberg, M., Spurkland, A., Jahnsen, T. (1993). Isolation and characterization of a human pseudogene for the regulatory subunit RIα of cAMP-dependent protein kinases and its sublocalization on chromosome 1. Genomics 15, 591–597.Google Scholar
  25. Tokunaga, K., Yoda, K., Sakiyama, S. (1985). Structure of a processed gene of mouse cytoplasmic γ-actin transposed into a BAM5 sequence: insertion has created 13 base-pair direct repeats. Nucleic Acids Res. 13, 3031–3042.Google Scholar
  26. Uhler, M., Chrivia, J., McKnight, G. (1986). Evidence for a second isoform of the catalytic subunit of cAMP-dependent protein kinase. J. Biol. Chem. 261, 15360–15363.Google Scholar
  27. Vanin, E. (1985). Processed pseudogenes: characteristics and evolution. Annu. Rev. Genet. 19, 253–272.Google Scholar
  28. Watson, J., Sutcliffe, J. (1987). Primate brain-specific cytoplasmic transcript of the Alu repeat family. Mol. Cell. Biol. 7, 3324–3327.Google Scholar
  29. Weiner, A., Deininger, P., Efstratiadis, A. (1986). Nonviral retroposons: genes, pseudogenes, and transposable elements generated by the reverse flow of genetic information. Annu. Rev. Biochem. 55, 631–661.Google Scholar
  30. Zakut-Houri, R., Oren, M., Bienz, B., Lavie, V., Hazum, S., Givol, D. (1983). A single gene and a pseudogene for the cellular tumour antigen p53. Nature 306, 594–597.Google Scholar

Copyright information

© Springer-Verlag New York Inc 1994

Authors and Affiliations

  • D. E. Cummings
    • 1
  • S. Edelhoff
    • 2
  • C. M. Disteche
    • 2
  • G. S. McKnight
    • 1
  1. 1.Department of Pharmacology, SJ-30University of WashingtonSeattleUSA
  2. 2.Department of PathologyUniversity of WashingtonSeattleUSA

Personalised recommendations