Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Oct 15;89(20):9885–9889. doi: 10.1073/pnas.89.20.9885

A protein methyltransferase specific for altered aspartyl residues is important in Escherichia coli stationary-phase survival and heat-shock resistance.

C Li 1, S Clarke 1
PMCID: PMC50238  PMID: 1409717

Abstract

Proteins are subject to spontaneous degradation reactions including the deamidation, isomerization, and racemization of asparaginyl and aspartyl residues. A major product of these reactions, the L-isoaspartyl residue, is recognized with high affinity by the protein-L-isoaspartate(D-aspartate) O-methyltransferase (EC 2.1.1.77). This enzyme catalyzes the methyl esterification of the L-isoaspartyl residue in a reaction that can initiate its conversion to the normal aspartyl configuration. To directly study the physiological role of this methyltransferase, especially with respect to the potential repair of isomerized aspartyl residues in aging proteins, we examined the ability of the bacterium Escherichia coli to survive in the absence of its activity. We utilized gene disruption techniques to replace the chromosomal copy of the pcm gene that encodes the methyltransferase with a kanamycin-resistance cassette to produce mutants that have no detectable L-isoaspartyl methyltransferase activity. Although no changes in exponential-phase growth were observed, pcm- mutants did not survive well upon extended culture into stationary phase or upon heat challenge at 55 degrees C. These results provide genetic evidence for a role of the L-isoaspartyl methyltransferase in the metabolism of altered proteins that can accumulate in aging cells and limit their viability.

Full text

PDF
9887

Images in this article

9887Image
on p.9887

Selected References

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

  1. Backman K., Chen Y. M., Magasanik B. Physical and genetic characterization of the glnA--glnG region of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3743–3747. doi: 10.1073/pnas.78.6.3743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barten D. M., O'Dea R. F. The function of protein carboxylmethyltransferase in eucaryotic cells. Life Sci. 1990;47(3):181–194. doi: 10.1016/0024-3205(90)90319-m. [DOI] [PubMed] [Google Scholar]
  3. Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene. 1982 Oct;19(3):327–336. doi: 10.1016/0378-1119(82)90023-3. [DOI] [PubMed] [Google Scholar]
  4. Clarke S. Protein carboxyl methyltransferases: two distinct classes of enzymes. Annu Rev Biochem. 1985;54:479–506. doi: 10.1146/annurev.bi.54.070185.002403. [DOI] [PubMed] [Google Scholar]
  5. Fu J. C., Ding L., Clarke S. Purification, gene cloning, and sequence analysis of an L-isoaspartyl protein carboxyl methyltransferase from Escherichia coli. J Biol Chem. 1991 Aug 5;266(22):14562–14572. [PubMed] [Google Scholar]
  6. Galletti P., Ciardiello A., Ingrosso D., Di Donato A., D'Alessio G. Repair of isopeptide bonds by protein carboxyl O-methyltransferase: seminal ribonuclease as a model system. Biochemistry. 1988 Mar 8;27(5):1752–1757. doi: 10.1021/bi00405a055. [DOI] [PubMed] [Google Scholar]
  7. Geiger T., Clarke S. Deamidation, isomerization, and racemization at asparaginyl and aspartyl residues in peptides. Succinimide-linked reactions that contribute to protein degradation. J Biol Chem. 1987 Jan 15;262(2):785–794. [PubMed] [Google Scholar]
  8. George-Nascimento C., Lowenson J., Borissenko M., Calderón M., Medina-Selby A., Kuo J., Clarke S., Randolph A. Replacement of a labile aspartyl residue increases the stability of human epidermal growth factor. Biochemistry. 1990 Oct 16;29(41):9584–9591. doi: 10.1021/bi00493a012. [DOI] [PubMed] [Google Scholar]
  9. Harding J. J., Beswick H. T., Ajiboye R., Huby R., Blakytny R., Rixon K. C. Non-enzymic post-translational modification of proteins in aging. A review. Mech Ageing Dev. 1989 Oct;50(1):7–16. doi: 10.1016/0047-6374(89)90054-7. [DOI] [PubMed] [Google Scholar]
  10. Ingrosso D., Fowler A. V., Bleibaum J., Clarke S. Sequence of the D-aspartyl/L-isoaspartyl protein methyltransferase from human erythrocytes. Common sequence motifs for protein, DNA, RNA, and small molecule S-adenosylmethionine-dependent methyltransferases. J Biol Chem. 1989 Nov 25;264(33):20131–20139. [PubMed] [Google Scholar]
  11. Jenkins D. E., Chaisson S. A., Matin A. Starvation-induced cross protection against osmotic challenge in Escherichia coli. J Bacteriol. 1990 May;172(5):2779–2781. doi: 10.1128/jb.172.5.2779-2781.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jenkins D. E., Schultz J. E., Matin A. Starvation-induced cross protection against heat or H2O2 challenge in Escherichia coli. J Bacteriol. 1988 Sep;170(9):3910–3914. doi: 10.1128/jb.170.9.3910-3914.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson B. A., Langmack E. L., Aswad D. W. Partial repair of deamidation-damaged calmodulin by protein carboxyl methyltransferase. J Biol Chem. 1987 Sep 5;262(25):12283–12287. [PubMed] [Google Scholar]
  14. Johnson B. A., Murray E. D., Jr, Clarke S., Glass D. B., Aswad D. W. Protein carboxyl methyltransferase facilitates conversion of atypical L-isoaspartyl peptides to normal L-aspartyl peptides. J Biol Chem. 1987 Apr 25;262(12):5622–5629. [PubMed] [Google Scholar]
  15. Johnson B. A., Ngo S. Q., Aswad D. W. Widespread phylogenetic distribution of a protein methyltransferase that modifies L-isoaspartyl residues. Biochem Int. 1991 Jul;24(5):841–847. [PubMed] [Google Scholar]
  16. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  17. Lange R., Hengge-Aronis R. Identification of a central regulator of stationary-phase gene expression in Escherichia coli. Mol Microbiol. 1991 Jan;5(1):49–59. doi: 10.1111/j.1365-2958.1991.tb01825.x. [DOI] [PubMed] [Google Scholar]
  18. Leyh T. S., Taylor J. C., Markham G. D. The sulfate activation locus of Escherichia coli K12: cloning, genetic, and enzymatic characterization. J Biol Chem. 1988 Feb 15;263(5):2409–2416. [PubMed] [Google Scholar]
  19. Li C., Clarke S. Distribution of an L-isoaspartyl protein methyltransferase in eubacteria. J Bacteriol. 1992 Jan;174(2):355–361. doi: 10.1128/jb.174.2.355-361.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Loewen P. C., Triggs B. L. Genetic mapping of katF, a locus that with katE affects the synthesis of a second catalase species in Escherichia coli. J Bacteriol. 1984 Nov;160(2):668–675. doi: 10.1128/jb.160.2.668-675.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lowenson J. D., Clarke S. Recognition of D-aspartyl residues in polypeptides by the erythrocyte L-isoaspartyl/D-aspartyl protein methyltransferase. Implications for the repair hypothesis. J Biol Chem. 1992 Mar 25;267(9):5985–5995. [PubMed] [Google Scholar]
  22. Lowenson J. D., Clarke S. Spontaneous degradation and enzymatic repair of aspartyl and asparaginyl residues in aging red cell proteins analyzed by computer simulation. Gerontology. 1991;37(1-3):128–151. doi: 10.1159/000213255. [DOI] [PubMed] [Google Scholar]
  23. Matin A., Auger E. A., Blum P. H., Schultz J. E. Genetic basis of starvation survival in nondifferentiating bacteria. Annu Rev Microbiol. 1989;43:293–316. doi: 10.1146/annurev.mi.43.100189.001453. [DOI] [PubMed] [Google Scholar]
  24. Matin A. The molecular basis of carbon-starvation-induced general resistance in Escherichia coli. Mol Microbiol. 1991 Jan;5(1):3–10. doi: 10.1111/j.1365-2958.1991.tb01819.x. [DOI] [PubMed] [Google Scholar]
  25. Mazodier P., Cossart P., Giraud E., Gasser F. Completion of the nucleotide sequence of the central region of Tn5 confirms the presence of three resistance genes. Nucleic Acids Res. 1985 Jan 11;13(1):195–205. doi: 10.1093/nar/13.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. McCann M. P., Kidwell J. P., Matin A. The putative sigma factor KatF has a central role in development of starvation-mediated general resistance in Escherichia coli. J Bacteriol. 1991 Jul;173(13):4188–4194. doi: 10.1128/jb.173.13.4188-4194.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. McFadden P. N., Clarke S. Conversion of isoaspartyl peptides to normal peptides: implications for the cellular repair of damaged proteins. Proc Natl Acad Sci U S A. 1987 May;84(9):2595–2599. doi: 10.1073/pnas.84.9.2595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McFadden P. N., Clarke S. Methylation at D-aspartyl residues in erythrocytes: possible step in the repair of aged membrane proteins. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2460–2464. doi: 10.1073/pnas.79.8.2460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mulvey M. R., Loewen P. C. Nucleotide sequence of katF of Escherichia coli suggests KatF protein is a novel sigma transcription factor. Nucleic Acids Res. 1989 Dec 11;17(23):9979–9991. doi: 10.1093/nar/17.23.9979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mulvey M. R., Sorby P. A., Triggs-Raine B. L., Loewen P. C. Cloning and physical characterization of katE and katF required for catalase HPII expression in Escherichia coli. Gene. 1988 Dec 20;73(2):337–345. doi: 10.1016/0378-1119(88)90498-2. [DOI] [PubMed] [Google Scholar]
  31. O'Connor C. M., Clarke S. Specific recognition of altered polypeptides by widely distributed methyltransferases. Biochem Biophys Res Commun. 1985 Nov 15;132(3):1144–1150. doi: 10.1016/0006-291x(85)91926-6. [DOI] [PubMed] [Google Scholar]
  32. O'Connor C. M., Germain B. J. Kinetic and electrophoretic analysis of transmethylation reactions in intact Xenopus laevis oocytes. J Biol Chem. 1987 Jul 25;262(21):10404–10411. [PubMed] [Google Scholar]
  33. Patel K., Borchardt R. T. Chemical pathways of peptide degradation. III. Effect of primary sequence on the pathways of deamidation of asparaginyl residues in hexapeptides. Pharm Res. 1990 Aug;7(8):787–793. doi: 10.1023/a:1015999012852. [DOI] [PubMed] [Google Scholar]
  34. Roszak D. B., Colwell R. R. Survival strategies of bacteria in the natural environment. Microbiol Rev. 1987 Sep;51(3):365–379. doi: 10.1128/mr.51.3.365-379.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Siegele D. A., Kolter R. Life after log. J Bacteriol. 1992 Jan;174(2):345–348. doi: 10.1128/jb.174.2.345-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stadtman E. R. Protein modification in aging. J Gerontol. 1988 Sep;43(5):B112–B120. doi: 10.1093/geronj/43.5.b112. [DOI] [PubMed] [Google Scholar]
  37. Winans S. C., Elledge S. J., Krueger J. H., Walker G. C. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. doi: 10.1128/jb.161.3.1219-1221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES