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. 1992 Jun;174(11):3782–3788. doi: 10.1128/jb.174.11.3782-3788.1992

Proline biosynthesis in Saccharomyces cerevisiae: analysis of the PRO3 gene, which encodes delta 1-pyrroline-5-carboxylate reductase.

M C Brandriss 1, D A Falvey 1
PMCID: PMC206069  PMID: 1592829

Abstract

The PRO3 gene of Saccharomyces cerevisiae encodes the 286-amino-acid protein delta 1-pyrroline-5-carboxylate reductase [L-proline:NAD(P+) 5-oxidoreductase; EC 1.5.1.2], which catalyzes the final step in proline biosynthesis. The protein has substantial similarity to the pyrroline carboxylate reductases of diverse bacterial species, soybean, and humans. Using RNA hybridization and measurements of enzyme activity, we have determined that the expression of the PRO3 gene appears to be constitutive. It is not repressed by the pathway end product (proline), induced by the initial substrate (glutamate), or regulated by the general control system. Its expression is not detectably altered when cells are grown in a wide range of nitrogen sources or when glycerol and ethanol replace glucose as the carbon source. The possibility that this enzyme has other functions in addition to proline biosynthesis is discussed.

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Selected References

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  1. Arst H. N., Jr, Jones S. A., Bailey C. R. A method for the selection of deletion mutations in the L-proline catabolism gene cluster of Aspergillus nidulans. Genet Res. 1981 Oct;38(2):171–195. doi: 10.1017/s0016672300020516. [DOI] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Botstein D., Falco S. C., Stewart S. E., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. doi: 10.1016/0378-1119(79)90004-0. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Brady R. A., Csonka L. N. Transcriptional regulation of the proC gene of Salmonella typhimurium. J Bacteriol. 1988 May;170(5):2379–2382. doi: 10.1128/jb.170.5.2379-2382.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brandriss M. C. Isolation and preliminary characterization of Saccharomyces cerevisiae proline auxotrophs. J Bacteriol. 1979 Jun;138(3):816–822. doi: 10.1128/jb.138.3.816-822.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brandriss M. C., Krzywicki K. A. Amino-terminal fragments of delta 1-pyrroline-5-carboxylate dehydrogenase direct beta-galactosidase to the mitochondrial matrix in Saccharomyces cerevisiae. Mol Cell Biol. 1986 Oct;6(10):3502–3512. doi: 10.1128/mcb.6.10.3502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brandriss M. C., Magasanik B. Genetics and physiology of proline utilization in Saccharomyces cerevisiae: enzyme induction by proline. J Bacteriol. 1979 Nov;140(2):498–503. doi: 10.1128/jb.140.2.498-503.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brandriss M. C., Magasanik B. Proline: an essential intermediate in arginine degradation in Saccharomyces cerevisiae. J Bacteriol. 1980 Sep;143(3):1403–1410. doi: 10.1128/jb.143.3.1403-1410.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Brandriss M. C., Magasanik B. Subcellular compartmentation in control of converging pathways for proline and arginine metabolism in Saccharomyces cerevisiae. J Bacteriol. 1981 Mar;145(3):1359–1364. doi: 10.1128/jb.145.3.1359-1364.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Brandriss M. C. Proline utilization in Saccharomyces cerevisiae: analysis of the cloned PUT2 gene. Mol Cell Biol. 1983 Oct;3(10):1846–1856. doi: 10.1128/mcb.3.10.1846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Delauney A. J., Verma D. P. A soybean gene encoding delta 1-pyrroline-5-carboxylate reductase was isolated by functional complementation in Escherichia coli and is found to be osmoregulated. Mol Gen Genet. 1990 May;221(3):299–305. doi: 10.1007/BF00259392. [DOI] [PubMed] [Google Scholar]
  14. Deutch A. H., Smith C. J., Rushlow K. E., Kretschmer P. J. Escherichia coli delta 1-pyrroline-5-carboxylate reductase: gene sequence, protein overproduction and purification. Nucleic Acids Res. 1982 Dec 11;10(23):7701–7714. doi: 10.1093/nar/10.23.7701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dougherty K. M., Brandriss M. C., Valle D. Cloning human pyrroline-5-carboxylate reductase cDNA by complementation in Saccharomyces cerevisiae. J Biol Chem. 1992 Jan 15;267(2):871–875. [PubMed] [Google Scholar]
  17. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  18. Gherardini F. C., Hobbs M. M., Stamm L. V., Bassford P. J., Jr Complementation of an Escherichia coli proC mutation by a gene cloned from Treponema pallidum. J Bacteriol. 1990 Jun;172(6):2996–3002. doi: 10.1128/jb.172.6.2996-3002.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hamilton P. T., Reeve J. N. Structure of genes and an insertion element in the methane producing archaebacterium Methanobrevibacter smithii. Mol Gen Genet. 1985;200(1):47–59. doi: 10.1007/BF00383311. [DOI] [PubMed] [Google Scholar]
  20. Hinnebusch A. G. Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae. Microbiol Rev. 1988 Jun;52(2):248–273. doi: 10.1128/mr.52.2.248-273.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kohl D. H., Schubert K. R., Carter M. B., Hagedorn C. H., Shearer G. Proline metabolism in N2-fixing root nodules: energy transfer and regulation of purine synthesis. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2036–2040. doi: 10.1073/pnas.85.7.2036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kozak M. Possible role of flanking nucleotides in recognition of the AUG initiator codon by eukaryotic ribosomes. Nucleic Acids Res. 1981 Oct 24;9(20):5233–5252. doi: 10.1093/nar/9.20.5233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lee D. C., Roeder R. G. Transcription of adenovirus type 2 genes in a cell-free system: apparent heterogeneity of initiation at some promoters. Mol Cell Biol. 1981 Jul;1(7):635–651. doi: 10.1128/mcb.1.7.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Matsuzawa T., Ishiguro I. Delta 1-pyrroline-5-carboxylate reductase from baker's yeast: further purification by affinity chromatography with 5' AMP-Sepharose 4B. Biochim Biophys Acta. 1980 Dec 4;616(2):381–383. doi: 10.1016/0005-2744(80)90155-2. [DOI] [PubMed] [Google Scholar]
  26. Matsuzawa T., Ishiguro I. delta 1-Pyrroline-5-carboxylate reductase from Baker's yeast. Purification, properties and its application in the assays of L-delta 1-pyrroline-5-carboxylate and L-ornithine in tissue. Biochim Biophys Acta. 1980 Jun 13;613(2):318–323. doi: 10.1016/0005-2744(80)90086-8. [DOI] [PubMed] [Google Scholar]
  27. Merrill M. J., Yeh G. C., Phang J. M. Purified human erythrocyte pyrroline-5-carboxylate reductase. Preferential oxidation of NADPH. J Biol Chem. 1989 Jun 5;264(16):9352–9358. [PubMed] [Google Scholar]
  28. Nagawa F., Fink G. R. The relationship between the "TATA" sequence and transcription initiation sites at the HIS4 gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8557–8561. doi: 10.1073/pnas.82.24.8557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Needleman R. B., Kaback D. B., Dubin R. A., Perkins E. L., Rosenberg N. G., Sutherland K. A., Forrest D. B., Michels C. A. MAL6 of Saccharomyces: a complex genetic locus containing three genes required for maltose fermentation. Proc Natl Acad Sci U S A. 1984 May;81(9):2811–2815. doi: 10.1073/pnas.81.9.2811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Phang J. M. The regulatory functions of proline and pyrroline-5-carboxylic acid. Curr Top Cell Regul. 1985;25:91–132. doi: 10.1016/b978-0-12-152825-6.50008-4. [DOI] [PubMed] [Google Scholar]
  32. Rose M., Botstein D. Structure and function of the yeast URA3 gene. Differentially regulated expression of hybrid beta-galactosidase from overlapping coding sequences in yeast. J Mol Biol. 1983 Nov 15;170(4):883–904. doi: 10.1016/s0022-2836(83)80193-4. [DOI] [PubMed] [Google Scholar]
  33. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Savioz A., Jeenes D. J., Kocher H. P., Haas D. Comparison of proC and other housekeeping genes of Pseudomonas aeruginosa with their counterparts in Escherichia coli. Gene. 1990 Jan 31;86(1):107–111. doi: 10.1016/0378-1119(90)90121-7. [DOI] [PubMed] [Google Scholar]
  35. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tomenchok D. M., Brandriss M. C. Gene-enzyme relationships in the proline biosynthetic pathway of Saccharomyces cerevisiae. J Bacteriol. 1987 Dec;169(12):5364–5372. doi: 10.1128/jb.169.12.5364-5372.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vogel H. J., Bonner D. M. ON THE GLUTAMATE-PROLINE-ORNITHINE INTERRELATION IN NEUROSPORA CRASSA. Proc Natl Acad Sci U S A. 1954 Aug;40(8):688–694. doi: 10.1073/pnas.40.8.688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wolfner M., Yep D., Messenguy F., Fink G. R. Integration of amino acid biosynthesis into the cell cycle of Saccharomyces cerevisiae. J Mol Biol. 1975 Aug 5;96(2):273–290. doi: 10.1016/0022-2836(75)90348-4. [DOI] [PubMed] [Google Scholar]
  39. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  40. Yeh G. C., Phang J. M. Stimulation of phosphoribosyl pyrophosphate and purine nucleotide production by pyrroline 5-carboxylate in human erythrocytes. J Biol Chem. 1988 Sep 15;263(26):13083–13089. [PubMed] [Google Scholar]
  41. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]

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