Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1987 Jan;83(1):85–91. doi: 10.1104/pp.83.1.85

Different Rates of Synthesis and Degradation of Two Chloroplastic Ammonium-Inducible NADP-Specific Glutamate Dehydrogenase Isoenzymes during Induction and Deinduction in Chlorella sorokiniana Cells 1

Newell F Bascomb 1,2, Donna E Prunkard 1,3, Robert R Schmidt 1
PMCID: PMC1056303  PMID: 16665221

Abstract

The kinetics of accumulation (per milliliter of culture) of the α- and β- subunits, associated with chloroplast-localized ammonium inducible nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase (NADP-GDH) isoenzymes, were measured during a 3 hour induction of synchronized daughter cells of Chlorella sorokiniana in 29 millimolar ammonium medium under photoautotrophic conditions. The β-subunit holoenzyme(s) accumulated in a linear manner for 3 hours without an apparent induction lag. A 40 minute induction lag preceded the accumulation of the α-subunit holoenzyme(s). After 120 minutes, the α-subunit ceased accumulating and thereafter remained at a constant level (i.e. steady state between synthesis and degradation). From pulsechase experiments, using 35SO4 and immunochemical procedures, the rate of synthesis of the α-subunit was shown to be greater than the β-subunit during the first 80 minutes of induction. The α- and β-subunits had different rates of degradation during the induction period (t½ = 50 versus 150 minutes, respectively) and during the deinduction period (t½ = 5 versus 13.5 minutes) after removal of ammonium from the culture. During deinduction, total NADP-GDH activity decreased with a half-time of 9 minutes. Cycloheximide completely inhibited the synthesis and degradation of both subunits. A model for regulation of expression of the NADP-GDH gene was proposed.

Full text

PDF
85

Selected References

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

  1. Ahmad I., Hellebust J. A. Nitrogen Metabolism of the Marine Microalga Chlorella autotrophica. Plant Physiol. 1984 Nov;76(3):658–663. doi: 10.1104/pp.76.3.658. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bascomb N. F., Schmidt R. R. Purification and Partial Kinetic and Physical Characterization of Two Chloroplast-Localized NADP-Specific Glutamate Dehydrogenase Isoenzymes and Their Preferential Accumulation in Chlorella sorokiniana Cells Cultured at Low or High Ammonium Levels. Plant Physiol. 1987 Jan;83(1):75–84. doi: 10.1104/pp.83.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bascomb N. F., Turner K. J., Schmidt R. R. Specific Polysome Immunoadsorption to Purify an Ammonium-Inducible Glutamate Dehydrogenase mRNA from Chlorella sorokiniana and Synthesis of Full Length Double-Stranded cDNA from the Purified mRNA. Plant Physiol. 1986 Jun;81(2):527–532. doi: 10.1104/pp.81.2.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bascomb N. F., Yeung A. T., Turner K. J., Schmidt R. R. Turnover of ammonium-inducible glutamate dehydrogenase during induction and its rapid inactivation after removal of inducer from Chlorella sorokiniana cells. J Bacteriol. 1981 Mar;145(3):1266–1272. doi: 10.1128/jb.145.3.1266-1272.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gronostajski R. M., Yeung A. T., Schmidt R. R. Purification and properties of the inducible nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase from Chlorella sorokiniana. J Bacteriol. 1978 May;134(2):621–628. doi: 10.1128/jb.134.2.621-628.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Israel D. W., Gronostajski R. M., Yeung A. T., Schmidt R. R. Regulation of accumulation and turnover of an inducible glutamate dehydrogenase in synchronous cultures of Chlorella. J Bacteriol. 1977 May;130(2):793–804. doi: 10.1128/jb.130.2.793-804.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Miller E. S., Brenchley J. E. Cloning and characterization of gdhA, the structural gene for glutamate dehydrogenase of Salmonella typhimurium. J Bacteriol. 1984 Jan;157(1):171–178. doi: 10.1128/jb.157.1.171-178.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Prunkard D. E., Bascomb N. F., Molin W. T., Schmidt R. R. Effect of Different Carbon Sources on the Ammonium Induction of Different Forms of NADP-Specific Glutamate Dehydrogenase in Chlorella sorokiniana Cells Cultured in the Light and Dark. Plant Physiol. 1986 Jun;81(2):413–422. doi: 10.1104/pp.81.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Prunkard D. E., Bascomb N. F., Robinson R. W., Schmidt R. R. Evidence for Chloroplastic Localization of an Ammonium-Inducible Glutamate Dehydrogenase and Synthesis of Its Subunit from a Cytosolic Precursor-Protein in Chlorella sorokiniana. Plant Physiol. 1986 Jun;81(2):349–355. doi: 10.1104/pp.81.2.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Schmidt R. R. Continuous dilution culture system for studies on gene-enzyme regulation in synchronous cultures of plant cells. In Vitro. 1974 Nov-Dec;10(5-6):306–320. doi: 10.1007/BF02615312. [DOI] [PubMed] [Google Scholar]
  12. Sitz T. O., Molloy G. R., Schmidt R. R. Evidence for turnover of ribulose-1,5-diphosphate carboxylase and continuous transcription of its structural gene throughout the cell cycle of the eucaryote Chlorella. Biochim Biophys Acta. 1973 Aug 10;319(1):103–108. doi: 10.1016/0005-2787(73)90045-2. [DOI] [PubMed] [Google Scholar]
  13. Talley D. J., White L. H., Schmidt R. R. Evidence for NADH- and NADPH-specific isozymes of glutamate dehydrogenase and the continuous inducibility of the NADPH-specific isozyme throughout the cell cycle of the eucaryote Chlorella. J Biol Chem. 1972 Dec 25;247(24):7927–7935. [PubMed] [Google Scholar]
  14. Toman P. D., Schmidt R. R. Comparison of Patterns of Accumulation of Ribulose Bisphosphate Carboxylase Antigen and Catalytic Activity and Measurement of Antigen Half-Life during the Cell Cycle of Chlorella sorokiniana. Plant Physiol. 1985 Nov;79(3):815–819. doi: 10.1104/pp.79.3.815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Turner K. J., Bascomb N. F., Lynch J. J., Molin W. T., Thurston C. F., Schmidt R. R. Evidence for messenger ribonucleic acid of an ammonium-inducible glutamate dehydrogenase and synthesis, covalent modification, and degradation of enzyme subunits in uninduced Chlorella sorokiniana cells. J Bacteriol. 1981 May;146(2):578–589. doi: 10.1128/jb.146.2.578-589.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Turner K. J., Gronostajski R. M., Schmidt R. R. Regulation of initial rate of induction of nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase during the cell cycle of synchronous Chlorella. J Bacteriol. 1978 Jun;134(3):1013–1019. doi: 10.1128/jb.134.3.1013-1019.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Vassef A. A., Flora J. B., Weeks J. G., Bibbs B. S., Schmidt R. R. The effects of enzyme synthesis and stability and of deoxyribonucleic acid replication on the cellular levels of aspartate transcarbamylase during the cell cycle of eucaryote Chlorella. J Biol Chem. 1973 Mar 25;248(6):1976–1985. [PubMed] [Google Scholar]
  18. Yeung A. T., Bascomb N. F., Turner K. J., Schmidt R. R. Regulation of accumulation of ammonium-inducible glutamate dehydrogenase catalytic activity and antigen during the cell cycle of fully induced, synchronous Chlorella sorokiniana cells. J Bacteriol. 1981 May;146(2):571–577. doi: 10.1128/jb.146.2.571-577.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yeung A. T., Turner K. J., Bascomb N. F., Schmidt R. R. Purification of an ammonium-inducible glutamate dehydrogenase and the use of its antigen affinity column-purified antibody in specific immunoprecipitation and immunoadsorption procedures. Anal Biochem. 1981 Jan 1;110(1):216–228. doi: 10.1016/0003-2697(81)90138-x. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES