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Profiles of nuclear and mitochondrial encoded mRNAs in developing and quiescent embryos of Artemia franciscana

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Abstract

Embryos of the brine shrimp Artemia franciscana are able to withstand long bouts of environmental anoxia by entering a quiescent state during which metabolism is greatly depressed. Recent evidence supports a global arrest of protein synthesis during quiescence. In this study we measured the amounts of mRNA for a mitochondrial-encoded subunit of cytochrome c oxidase (COX I) and for nuclear-encoded actin during aerobic development, anaerobiosis, and aerobic acidosis (artificial quiescence imposed by intracellular acidification under aerobic conditions). The levels of both COX I and actin transcripts increased significantly during aerobic development. COX I mRNA levels were tightly correlated with previous measures of COX catalytic activity, which suggests that COX synthesis could be regulated by message concentration during aerobic development. The ontogenetic increase for these mRNAs was blocked by anoxia and aerobic acidosis. Importantly, the levels of COX I and actin mRNA did not decline appreciably during the 6 h bouts of quiescence, even though protein synthesis is acutely arrested by these same treatments. Thus, the constancy of mRNA levels during quiescence indicate that reduced protein synthesis is not caused by message limitation, but rather, is likely controlled at the translational level. One advantage of this regulatory mechanism is the conservation of mRNA molecules during quiescence, which would potentially favor a quick resumption of translation as soon as oxygen is returned to the embryos. Finally, because anoxia and aerobic acidosis are both characterized by acidic intracellular pH, the reduction in pH may serve, directly or indirectly, as one signal regulating levels of mRNA in this embryo during quiescence.

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References

  1. Hand SC, Gnaiger E: Anaerobic dormancy quantified in Artemia embryos: A calorimetric test of the control mechanism. Science 239: 1425–1427, 1988

    Google Scholar 

  2. Busa WB, Crowe JH: Intracellular pH regulates transitions between dormancy and development of the brine shrimp (Artemia salina) embryos. Science 221: 366–368, 1983

    Google Scholar 

  3. Hofmann GE, Hand SC: Arrest of cytochrome-c oxidase synthesis coordinated with catabolic arrest in dormant Artemia embryos. Am J Physiol 27: R1184-R1191, 1990

    Google Scholar 

  4. Hofmann GE, Hand SC: Global arrest of translation during invertebrate quiescence. Proc Natl Acad Sci USA 91: 8492–8496, 1994

    Google Scholar 

  5. Clegg JS, Conte FP: Cellular and developmental biology of Artemia. In: G Persoone, P Sorgeloos, O Roels, E Jaspers (eds). The brine shrimp Artemia. Universa, Whetteren, Belgium, vol 2 Physiology, biochemistry and molecular biology, pp 83–103, 1980

    Google Scholar 

  6. Hand SC: Metabolic dormancy in aquatic invertebrates. Adv Comp Environ Physiol 8: 1–49, 1991

    Google Scholar 

  7. Clegg JS, Golub AL: Protein synthesis in Artemia salina embryos. II. Resumption of RNA and protein synthesis upon cessation of dormancy in encysted gastrulae. Dev Biol 19: 178–200, 1969

    Google Scholar 

  8. Clegg JS: Protein synthesis in the absence of cell division during the development of Artemia salina embryos. Nature 212: 517–519, 1966

    Google Scholar 

  9. Hofmann GE, Hand SC: Subcellular differentiation arrested in Artemia embryos under anoxia: evidence supporting a regulatory role for intracellular pH. J Exp Zool 253: 287–302, 1990

    Google Scholar 

  10. Anchordoguy TJ, Hofmann GE, Hand, SC: Extension of enzyme halflife during quiescence in Artemia embryos. Am J Physiol 264: R85-R89, 1993

    Google Scholar 

  11. Busa WB, Crowe JH, Matson GB: Intracellular pH and the metabolic status of dormant and developing Artemia embryos. Arch Biochem Biophys 216: 711–718, 1982

    Google Scholar 

  12. Kwast KE, Shapiro JI, Rees BB, Hand SC: Oxidative phosphorylation and the realkalinization of intracellular pH during recovery from anoxia in Artemia franciscana embryos. Biochim Biophys Acta 1232: 5–12, 1995

    Google Scholar 

  13. Carpenter JF, Hand SC: Arrestment of carbohydrate metabolism during anaerobic dormancy and aerobic acidosis in Artemia embryos: determination of control points. J Comp Physiol 156: 451–459, 1986

    Google Scholar 

  14. Rees BB, Ropson I, Hand SC: Kinetic properties of hexokinase under near physiological conditions: Relation to metabolic arrest in Artemia embryos during anoxia. J Biol Chem 264: 15410–15417, 1989

    Google Scholar 

  15. Anchordoguy TJ, Hand SC: Acute blockage of the ubiquitin-mediated proteolytic pathway during invertebrate quiescence. Am J Physiol 267: R895-R900, 1994

    Google Scholar 

  16. Poyton RO: Cooperative interaction between mitochondrial and nuclear genomes: cytochrome c oxidase assembly as a model. Curr Top Cell Regul 17:231–295, 1980

    Google Scholar 

  17. Kwast KE, Hand SC: Oxygen and pH regulation of protein synthesis in mitochondria from Artemia franciscana embryos. Biochem J 313: 207–213, 1996

    Google Scholar 

  18. Kwast K, Hand SC: Regulatory features of protein synthesis in isolated mitochondria from Artemia embryos. Amer J Physiol 265: R1238-R1246, 1993

    Google Scholar 

  19. Capaldi RA: Structure and function of cytochrome c oxidase. Annu Rev Biochem 59: 569–596, 1990

    Google Scholar 

  20. Gould SJ, Subramani S, Scheffler IE: Use of DNA polymerase chain reaction for homology probing: Isolation of partial cDNA or genomic clones encoding the iron-sulfur protein of succinate dehydrogenase from several species. Proc Natl Acad Sci USA 86: 1934–1938, 1989

    Google Scholar 

  21. Valverde JR, Batuecas B, Garesse R: The complete mitochondrial sequence of the crustacean Artemia franciscana. J Mol Evol 39: 400–408

  22. Crawford DL: Nuclear genes for the copepod Calanus finmarchicus. Mol Mar Biol Biotech 4: 241–247, 1995

    Google Scholar 

  23. Macias M-T, Sastre L: Molecular cloning and expression of four actin isoforms during Artemia development. Nucl Acids Res 18(17): 5219–5225, 1990

    Google Scholar 

  24. Anchordoguy TJ, Crawford DL, Hardewig I, Hand, SC: Heterogeneity of DNA binding to membranes used in quantitative dot blots. BioTechniques, 1995 (in press)

  25. Koller HT, Frondorf KA, Maschner PD, Vaughn JC: In vitro transcription from multiple spacer rRNA gene promotors during early development and evolution of the intergenic spacer in the brine shrimp Artemia. Nucl Acids Res 15(13): 5391–5411, 1987

    Google Scholar 

  26. McClean DK, Warner AH: Aspects of nucleic acid metabolism during development of the brine shrimp Artemia salina. Dev Biol 24: 88–105, 1971

    Google Scholar 

  27. Hofmann GE, Hand SC: Comparison of messenger RNA pools in active and dormant Artemia franciscana embryos: evidence for translational control. J Exp Biol 164: 103–116, 1992

    Google Scholar 

  28. Hand SC, Hardewig I: Downregulation of cellular metabolism during environmental stress: Mechanisms and implications. Annu Rev Physiol 58:539–563, 1996

    Google Scholar 

  29. Clegg JS: Metabolic studies of cryptobiosis in encysted embryos of Artemia salina. Comp Biochem Physiol 20: 801–809, 1967

    Google Scholar 

  30. Slegers H, De Herdt E, Kondo M: Non-polysomal poly(A)-containing messenger ribonucleoproteins of cryptobiotic gastrulae of Artemia salina. Eur J Biochem 117: 111–120, 1981

    Google Scholar 

  31. Diaz-Guerra M, Quintanilla M, Palermo I, Sastre L, Renart J: Differential expression of a gene highly homologous tp c-ras during the development of the brine shrimp Artemia. Biochem Biophys Res Comm 162(2): 802–808, 1989

    Google Scholar 

  32. Escalante R, Garcia-Saez A, Ortega M-A, Sastre L: Gene expression after resumption of development of Artemia franciscana cryptobiotic embryos. Biochem Cell Biol 72: 78–83, 1994

    Google Scholar 

  33. Hood D: Co-ordinate expression of cytochrome c oxidase subunit III and VIc mRNAs in rat tissues. Biochem J 269: 503–506, 1990

    Google Scholar 

  34. Gagnon J, Kurowski TT, Wiesner RJ, Zak R: Correlations between a nuclear and mitochondrial mRNA of cytochrome c oxidase subunits, enzymatic activity and total mRNA content, in rat tissues. Mol Cel Biochem 107: 21–29, 1991

    Google Scholar 

  35. Hood DA, Zak R, Pette D: Chronic stimulation of rat skelatal muscle induces coordinate increases in mitochondrial and nuclear mRNAs of cytochrome-c-oxidase subunits. Eur J Biochem 179: 275–280, 1989

    Google Scholar 

  36. Costanzo MC, Fox TD: Control of mitochondrial gene expression in Saccharomyces cerevisiae. Annu Rev Genet 24: 91–113, 1990

    Google Scholar 

  37. Fox TD, Shen Z: Positive control of translation in organellar genetic systems. In: AJP Brown, MF Tuite, JEG McCarthy (eds) Protein Synthesis and Targeting in Yeast. Springer-Verlag, Berlin, pp 157–166, 1993

    Google Scholar 

  38. Kloekner-Gruissem B, McEwan JM, Poyton RO: Nuclear functions required for cytochrome c oxidase biogenesis in Saccharomyces cerevisiae: multiple trans-acting nuclear genes exert specific effects on expression of each of the cytochrome c oxidase subunits encoded in mitochondrial DNA. Curr Genet 12: 311–22, 1987

    Google Scholar 

  39. Tzagoloff A, Dieckmann CL: PET genes of Saccharomyces cerevisiae. Microbiol Rev 54: 211–25, 1990

    Google Scholar 

  40. Surdej P, Riedl A, Jacobs-Lorena M: Regulation of mRNA stability in development. Annu Rev Genet 28: 263–282, 1994

    Google Scholar 

  41. Grosfeld H, Littauer UZ: The translation in vitro of mRNA from developing cysts of Artemia salina. Eur J Biochem 70: 589–599, 1976

    Google Scholar 

  42. Aviram M, Hershko A: Influence of ATP depletion on the degradation of rapidly labeled RNA in cultured hepatoma cells. Biochem Biophys Res Comm 65(4): 1303–1310, 1975

    Google Scholar 

  43. Fan DP, Higa A, Levinthal C: Messenger RNA decay and protection. J Mol Biol 8: 210–222, 1964

    Google Scholar 

  44. Georgellis D, Barlow T, Arvidson S, van Gabain A: Retarded RNA turnover in Escherichia coli: a means of maintaining gene expression during anaerobiosis. Mol Microbiol 9(2): 375–381, 1993

    Google Scholar 

  45. Lefebvre VHL, Van Steenbrugge M, Beckers V, Roberfroid M, Buc-Calderon P: Adenine nucleotides and inhibition of protein synthesis in isolated hepatocytes incubated under different Po2 levels. Arch Biochem Biophys 304(2): 322–331,1993

    Google Scholar 

  46. Kadowaki T, Kitawaga Y. Hypoxic depression of mitochondrial mRNA levels in HeLa cells. Exp Cell Res 192: 243–247, 1991

    Google Scholar 

  47. Conaway RC, Conaway JW: General initiation factors for RNA polymerase II. Annu Rev Biochem 62: 161–190, 1993

    Google Scholar 

  48. Hofmann GE: Aspects of development and the regulation of protein synthesis in dormant embryos of the brine shrimp, Artemia franciscana. Ph.D. Dissertation, University of Colorado, Boulder, 1992

    Google Scholar 

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Author notes

  1. Iris Hardewig

    Present address: Alfred-Wegener-Institute für Polar-und Meeresforschung, 27515, Columbusstrasse, Bremerhaven, Germany

Authors and Affiliations

  1. Department of EPO Biology, University of Colorado, 80309-0334, Boulder, CO

    Iris Hardewig, Thomas J. Anchordoguy & Steven C. Hand

  2. Department of Organismal Biology and Anatomy, University of Chicago, 60637, Chicago, IL, USA

    Douglas L. Crawford

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  1. Iris Hardewig
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  2. Thomas J. Anchordoguy
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  3. Douglas L. Crawford
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  4. Steven C. Hand
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Hardewig, I., Anchordoguy, T.J., Crawford, D.L. et al. Profiles of nuclear and mitochondrial encoded mRNAs in developing and quiescent embryos of Artemia franciscana . Mol Cell Biochem 158, 139–147 (1996). https://doi.org/10.1007/BF00225840

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  • DOI: https://doi.org/10.1007/BF00225840

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