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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1990 Dec;81(12):1272–1280. doi: 10.1111/j.1349-7006.1990.tb02690.x

Identification of Members of the Protein Phosphatase 1 Gene Family in the Rat and Enhanced Expression of Protein Phosphatase 1α Gene in Rat Hepatocellular Carcinomas

Kazunori Sasaki 1, Hiroshi Shima 1, Yoshinori Kitagawa 1, Shozo Irino 2, Takashi Sugimura 1, Minako Nagao 1,✉
PMCID: PMC5918009  PMID: 2177460

Abstract

We isolated four kinds of cDNA clones of isotypes of catalytic subunits of protein phosphatase 1 (PP‐1) from rat liver and testis cDNA libraries. For the cloning, cDNA fragments of dis2ml and dis2m2, which encode mouse PP‐1 catalytic subunits, were used as probes. Two of the four isotypes were thought to be derived from the same gene and produced by alternative splicing. Based on the comparative study of their nucleotide and deduced amino acid sequences with those reported, these cDNA clones were named rat PP‐1α, PP‐1γ1, PP‐1γ2 and PP‐δ. The deduced amino acid sequences of these four cDNA clones showed about 90% identity. Their amino‐terminal regions were highly conserved, and their differences were mainly in the carboxy‐terminal regions. Furthermore, several amino acids located in the middle regions of the peptides were conserved in all the isotypes of the catalytic subunits of PP‐1, PP‐2A, PP‐2B and PP‐2C. These conserved regions are suggested to be the functional domains of the catalytic subunits of protein phosphatases. Rat hepatocellular carcinomas induced by a food mutagen, 2‐amino‐3,8‐dimethylimidazo[4,5‐f]quinoxaline showed increased expression of PP‐1α, but no increased expression of PP‐1γ1, PP‐1γ2 or PP‐1δ. Involvement of PP‐1α in hepatocarcinogenesis or in hepatic cell proliferation was suspected.

Keywords: Protein phosphatase 1, Gene family, Alternative splicing, Conserved amino acid

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The nucleotide sequence data reported in this paper will appear in the DDBJ, EMBL and Gene Bank Nucleotide Sequence Database under the accession number D90163‐D90166

REFERENCES

  • 1. ) Ingebritsen , T. S. and Cohen , P.The protein phosphatases involved in cellular regulation . Eur. J. Biochem. , 132 , 255 – 261 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Ingebritsen , T. S. and Cohen , P.Protein phosphatases: properties and role in cellular regulation . Science , 221 , 331 – 339 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Cohen , P.The structure and regulation of protein phosphatases . Annu. Rev. Biochem. , 58 , 453 – 508 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 4. ) Ballou , L. M. , Brautigan , D. L. and Fischer , E. H.Subunit structure and activation of inactive phosphorylase phosphatase . Biochemistry , 22 , 3393 – 3399 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Hubbard , M. J. and Cohen , P.The glycogen‐binding subunit of protein phosphatase 1G from rabbit skeletal muscle: further characterisation of its structure and glycogen‐binding properties . Eur. J. Biochem. , 180 , 457 – 465 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Berndt , N. , Campbell , D. G. , Caudwell , F. B. , Cohen , P. , da Cruz e Silva , E. F. , da Cruz e Silva , O.B. and Cohen , P. T. W.Isolation and sequence analysis of a cDNA clone encoding a type‐1 protein phosphatase catalytic subunit: homology with protein phosphatase 2A . FEBS Lett. , 223 , 340 – 346 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Bai , G. , Zhang , Z. , Amin , J. , Deans‐Zirattu , S. A. and Lee , E. Y. C.Molecular cloning of a cDNA for the catalytic subunit of rabbit muscle phosphorylase phosphatase . FASEB J. , 2 , 3010 – 3016 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Cohen , P. T. W.Two isoforms of protein phosphatase 1 may be produced from the same gene . FEBS Lett. , 232 , 17 – 23 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Ohkura , H. , Kinoshita , N. , Miyatani , S. , Toda , T. and Yanagida , M.The fission yeast dis2+ gene required for chromosome disjoining encodes one of two putative type 1 protein phosphatases . Cell , 57 , 997 – 1007 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) da Cruz e Silva , O. B. , Alemany , S. , Campbell , D. G. and Cohen , P. T. W.Isolation and sequence analysis of a cDNA clone encoding the entire catalytic subunit of a type‐2A protein phosphatase . FEBS Lett. , 221 , 415 – 422 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) da Cruz e Silva , O. B. and Cohen , P. T. W.A second catalytic subunit of type‐2A protein phosphatase from rabbit skeletal muscle . FEBS Lett. , 226 , 176 – 178 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Kitagawa , Y. , Tahira , T. , Ikeda , I. , Kikuchi , K. , Tsuiki , S. , Sugimura , T. and Nagao , M.Molecular cloning of cDNA for the catalytic subunit of rat liver type 2A protein phosphatase, and detection of high levels of expression of the gene in normal and cancer cells . Biochim. Biophys. Acta , 951 , 123 – 129 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Kitagawa , Y. , Sakai , R. , Tahira , T. , Tsuda , H. , Ito , N. , Sugimura , T. and Nagao , M.Molecular cloning of rat phosphoprotein phosphatase 2Aβ cDNA and increased expressions of phosphatase 2Aα and 2Aβ in rat liver tumors . Biochem. Biophys. Res. Commun. , 157 , 821 – 827 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Ito , A. , Hashimoto , T. , Hirai , M. , Takeda , T. , Shuntoh , H. , Kuno , T. and Tanaka , C.The complete primary structure of calcineurin A, a calmodulin binding protein homologous with protein phosphatases 1 and 2A . Biochem. Biophys. Res. Commun. , 163 , 1492 – 1497 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Kuno , T. , Takeda , T. , Hirai , M. , Ito , A. , Mukai , H. and Tanaka , C.Evidence for a second isoform of the catalytic subunit of calmodulin‐dependent protein phosphatase (calcineurin A) . Biochem. Biophys. Res. Commun. , 165 , 1352 – 1358 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) McGowan , C. H. and Cohen , P.Identification of two isoenzymes of protein phosphatase 2C in both rabbit skeletal muscle and liver . Eur. J. Biochem. , 166 , 713 – 722 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) McGowan , C. H. , Campbell , D. G. and Cohen , P.Primary structure analysis proves that protein phosphatases 2C1 and 2C2 are isozymes . Biochim. Biophys. Acta , 930 , 279 – 282 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Cohen , P. T. W. , Brewis , N. D. , Hughes , V. and Mann , D. J.Protein serine/threonine phosphatase; an expanding family . FEBS Lett. , 268 , 355 – 359 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) da Cruz e Silva , O. B. , da Cruz e Silva , E. F. and Cohen , P. T. W.Identification of a novel protein phosphatase catalytic subunit by cDNA cloning . FEBS Lett. , 242 , 106 – 110 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Dombrádi , V. , Axton , J. M. , Glover , D. M. and Cohen , P. T. W.Molecular cloning and chromosomal localization of a novel Drosophila protein phosphatase . FEBS Lett. , 247 , 391 – 395 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Sakai , R. , Ikeda , I. , Kitani , H. , Fujiki , H. , Takaku , F. , Rapp , U. , Sugimura , T. and Nagao , M.Flat reversion by okadaic acid of raf and ret‐II transformants . Proc. Natl. Acad. Sci. USA , 86 , 9946 – 9950 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. ) Shineha , R. , Kikuchi , K. , Tamura , S. , Hiraga , A. , Suzuki , Y. and Tsuiki , S.Particulate‐associated protein phosphatases of rat hepatomas as compared with the enzymes of rat liver . Jpn. J. Cancer Res. , 81 , 161 – 168 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Gubler , U. and Hoffman , B. J.A simple and very efficient method for generating cDNA libraries . Gene , 25 , 263 – 269 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Hyunh , T. V. , Young , R. A. and Davis , R. W. “ DNA Cloning: A Practical Approach ,” Vol. I , ed. Glover D. , pp. 49 – 78 ( 1985. ). IRL Press; , Oxford . [Google Scholar]
  • 25. ) Feinberg , A. P. and Vogelstein , B.A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity . Anal. Biochem. , 132 , 6 – 13 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Chomczynski , P. and Sacchi , N.Single‐step method of RNA isolation by acid guanidinium thiocyanate‐phenolchloroform extraction . Anal. Biochem. , 162 , 156 – 159 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 27. ) Maniatis , T. , Fritsch , E. F. and Sambrook , J. “ Molecular Cloning: A Laboratory Manual ” ( 1982. ). Cold Spring Harbor Laboratory; , New York . [Google Scholar]
  • 28. ) Kitagawa , Y. , Sasaki , K. , Shima , H. , Shibuya , M. , Sugimura , T. and Nagao , M.Protein phosphatases possibly involved in rat spermatogenesis . Biochem. Biophys. Res. Commun. , 170 , 230 – 235 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Sasaki , K. , Kitagawa , Y. , Shima , H. , Irino , S. , Sugimura , T. and Nagao , M.Production of shorter mRNA for protein phosphatase 2Aβ by alternative poly(A) addition . Biochem. Biophys. Res. Commun. , 170 , 169 – 175 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Sandberg , M. , Skålhegg , B. and Jahnsen , T.The two mRNA forms for the type Iα regulatory subunit of cAMP‐dependent protein kinase from human testis are due to the use of different polyadenylation site signals . Biochem. Biophys. Res. Commun. , 167 , 323 – 330 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 31. ) Dobner , P. R. , Kislauskis , E. , Wentworth , B. M. and Villa‐Komaroff , L.Alternative 5′ exons either provide or deny an initiator methionine codon to the same α‐tubulin coding region . Nucleic Acid Res. , 15 , 199 – 218 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. ) Tamura , S. , Lynch , K. R. , Larner , J. , Fox , J. , Yasui , A. , Kikuchi , K. , Suzuki , Y. and Tsuiki , S.Molecular cloning of rat type 2C (IA) protein phosphatase mRNA . Proc. Natl. Acad. Sci. USA , 86 , 1796 – 1800 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. ) Thiede , M. A. , Yoon , K. , Golub , E. E. , Noda , M. and Rodan , G. A.Structure and expression of rat osteosarcoma (ROS 17/2.8) alkaline phosphatase: product of a single copy gene . Proc. Natl. Acad. Sci. USA , 85 , 319 – 323 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. ) Charbonneau , H. , Tonks , N. K. , Kumar , S. , Diltz , C. D. , Harrylock , M. , Cool , D. E. , Krebs , E. G. , Fischer , E. H. and Walsh , K. A.Human placenta protein‐tyrosine‐phosphatase: amino acid sequence and relationship to a family of receptor‐like proteins . Proc. Natl. Acad. Sci. USA , 86 , 5252 – 5256 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35. ) Cool , D. E. , Tonks , N. K. , Charbonneau , H. , Walsh , K. A. , Fisher , E. H. and Krebs , E. G.cDNA isolates from a human T‐cell library encodes a member of the protein‐tyrosine‐phosphatase family . Proc. Natl. Acad. Sci. USA , 86 , 5257 – 5261 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36. ) Booher , R. and Beach , D.Involvement of a type 1 protein phosphatase encoded by bws1+ in fission yeast mitotic control . Cell , 57 , 1009 – 1016 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 37. ) Dombrádi , V. , Axton , J. M. , Glover , D. M. and Cohen , P. T. W.Cloning and chromosomal localization of Drosophila cDNA encoding the catalytic subunit of protein phosphatase 1α: high conservation between mammalian and insect sequences . Eur. J. Biochem. , 183 , 603 – 610 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 38. ) Doonan , J. H. and Morris , N. R.The bimG gene of Aspergillus nidulans, required for completion of anaphase, encodes a homolog of mammalian phosphoprotein phosphatase 1 . Cell , 57 , 987 – 996 ( 1989. ). [DOI] [PubMed] [Google Scholar]

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