Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1990 Mar 15;266(3):901–907.

Characterization of the mRNAs encoding the gene 2 sarcoplasmic/endoplasmic-reticulum Ca2+ pump in pig smooth muscle.

J A Eggermont 1, F Wuytack 1, R Casteels 1
PMCID: PMC1131224  PMID: 2158303

Abstract

The gene 2 sarcoplasmic/endoplasmic-reticulum (SR/ER) Ca2+ pump is expressed in slow skeletal and cardiac muscle, smooth muscle and non-muscle tissues. We have analysed the gene 2 Ca2+ pump mRNAs using a panel of anti-sense RNA probes which recognize either the muscle (class 1) or the non-muscle (class 2) transcript, or both. In pig smooth muscle, we confirmed the presence of the class 1 and class 2 mRNAs of 4.4 kb length and we also detected a third mRNA of 8.0 kb which reacted with both the class 1 and class 2 riboprobes. A 4.2 kb cDNA corresponding to the 3' part of the 8.0 kb mRNA was cloned from a pig gastric smooth muscle cDNA library. Nucleotide sequence analysis of this clone revealed that the 8.0 kb mRNA (class 3 transcript) contained both the non-muscle-specific and the muscle-specific exons separated by a 2.4 kb intron which has not been removed. The class 3-mRNA-encoded SR/ER Ca2+ pump is identical to the class 2-encoded non-muscle isoform. Northern blot analysis demonstrated that, in cardiac muscle, the class 1 mRNA (encoding the muscle isoform) is the predominant messenger, whereas in non-muscle tissues the class 2 and 3 mRNAs (encoding the non-muscle isoform) predominate. In smooth muscle all three mRNA types are present. The tissue distribution of the mRNA types suggests a tissue-dependent processing of the primary transcript of the sarcoplasmic/endoplasmic reticulum Ca2+ pump gene 2.

Full text

PDF
901

Images in this article

902Fig. 1.
on p.902
905Fig. 4.
on p.905

Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandl C. J., Green N. M., Korczak B., MacLennan D. H. Two Ca2+ ATPase genes: homologies and mechanistic implications of deduced amino acid sequences. Cell. 1986 Feb 28;44(4):597–607. doi: 10.1016/0092-8674(86)90269-2. [DOI] [PubMed] [Google Scholar]
  3. Brandl C. J., deLeon S., Martin D. R., MacLennan D. H. Adult forms of the Ca2+ATPase of sarcoplasmic reticulum. Expression in developing skeletal muscle. J Biol Chem. 1987 Mar 15;262(8):3768–3774. [PubMed] [Google Scholar]
  4. Burk S. E., Lytton J., MacLennan D. H., Shull G. E. cDNA cloning, functional expression, and mRNA tissue distribution of a third organellar Ca2+ pump. J Biol Chem. 1989 Nov 5;264(31):18561–18568. [PubMed] [Google Scholar]
  5. Carmichael G. G., McMaster G. K. The analysis of nucleic acids in gels using glyoxal and acridine orange. Methods Enzymol. 1980;65(1):380–391. doi: 10.1016/s0076-6879(80)65049-6. [DOI] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Eggermont J. A., Wuytack F., De Jaegere S., Nelles L., Casteels R. Evidence for two isoforms of the endoplasmic-reticulum Ca2+ pump in pig smooth muscle. Biochem J. 1989 Jun 15;260(3):757–761. doi: 10.1042/bj2600757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gunteski-Hamblin A. M., Greeb J., Shull G. E. A novel Ca2+ pump expressed in brain, kidney, and stomach is encoded by an alternative transcript of the slow-twitch muscle sarcoplasmic reticulum Ca-ATPase gene. Identification of cDNAs encoding Ca2+ and other cation-transporting ATPases using an oligonucleotide probe derived from the ATP-binding site. J Biol Chem. 1988 Oct 15;263(29):15032–15040. [PubMed] [Google Scholar]
  9. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  10. Komuro I., Kurabayashi M., Shibazaki Y., Takaku F., Yazaki Y. Molecular cloning and characterization of a Ca2+ + Mg2+-dependent adenosine triphosphatase from rat cardiac sarcoplasmic reticulum. Regulation of its expression by pressure overload and developmental stage. J Clin Invest. 1989 Apr;83(4):1102–1108. doi: 10.1172/JCI113989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Korczak B., Zarain-Herzberg A., Brandl C. J., Ingles C. J., Green N. M., MacLennan D. H. Structure of the rabbit fast-twitch skeletal muscle Ca2+-ATPase gene. J Biol Chem. 1988 Apr 5;263(10):4813–4819. [PubMed] [Google Scholar]
  12. 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]
  13. Lompre A. M., de la Bastie D., Boheler K. R., Schwartz K. Characterization and expression of the rat heart sarcoplasmic reticulum Ca2+-ATPase mRNA. FEBS Lett. 1989 May 22;249(1):35–41. doi: 10.1016/0014-5793(89)80010-9. [DOI] [PubMed] [Google Scholar]
  14. Lytton J., MacLennan D. H. Molecular cloning of cDNAs from human kidney coding for two alternatively spliced products of the cardiac Ca2+-ATPase gene. J Biol Chem. 1988 Oct 15;263(29):15024–15031. [PubMed] [Google Scholar]
  15. Lytton J., Zarain-Herzberg A., Periasamy M., MacLennan D. H. Molecular cloning of the mammalian smooth muscle sarco(endo)plasmic reticulum Ca2+-ATPase. J Biol Chem. 1989 Apr 25;264(12):7059–7065. [PubMed] [Google Scholar]
  16. MacLennan D. H., Brandl C. J., Korczak B., Green N. M. Amino-acid sequence of a Ca2+ + Mg2+-dependent ATPase from rabbit muscle sarcoplasmic reticulum, deduced from its complementary DNA sequence. Nature. 1985 Aug 22;316(6030):696–700. doi: 10.1038/316696a0. [DOI] [PubMed] [Google Scholar]
  17. Mierendorf R. C., Pfeffer D. Direct sequencing of denatured plasmid DNA. Methods Enzymol. 1987;152:556–562. doi: 10.1016/0076-6879(87)52061-4. [DOI] [PubMed] [Google Scholar]
  18. Rasmussen H., Barrett P. Q. Calcium messenger system: an integrated view. Physiol Rev. 1984 Jul;64(3):938–984. doi: 10.1152/physrev.1984.64.3.938. [DOI] [PubMed] [Google Scholar]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. de la Bastie D., Wisnewsky C., Schwartz K., Lompré A. M. (Ca2+ + Mg2+)-dependent ATPase mRNA from smooth muscle sarcoplasmic reticulum differs from that in cardiac and fast skeletal muscles. FEBS Lett. 1988 Feb 29;229(1):45–48. doi: 10.1016/0014-5793(88)80794-4. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES