Advertisement

Planta

, Volume 221, Issue 5, pp 619–636 | Cite as

Gene structure and molecular analysis of the laccase-like multicopper oxidase (LMCO) gene family in Arabidopsis thaliana

  • Bonnie C. McCaig
  • Richard B. Meagher
  • Jeffrey F. D. DeanEmail author
Original Article

Abstract

Completed genome sequences have made it clear that multicopper oxidases related to laccase are widely distributed as multigene families in higher plants. Laccase-like multicopper oxidase (LMCO) sequences culled from GenBank and the Arabidopsis thaliana genome, as well as those from several newly cloned genes, were used to construct a gene phylogeny that clearly divided plant LMCOs into six distinct classes, at least three of which predate the evolutionary divergence of angiosperms and gymnosperms. Alignments of the predicted amino acid sequences highlighted regions of variable sequence flanked by the highly conserved copper-binding domains that characterize members of this enzyme family. All of the predicted proteins contained apparent signal sequences. The expression of 13 of the 17 LMCO genes in A. thaliana was assessed in different tissues at various stages of development using RT-PCR. A diversity of expression patterns was demonstrated with some genes being expressed in a constitutive fashion, while others were only expressed in specific tissues at a particular stage of development. Only a few of the LMCO genes were expressed in a pattern that could be considered consistent with a major role for these enzymes in lignin deposition. These results are discussed in the context of other potential physiological functions for plant LMCOs, such as iron metabolism and wound healing.

Keywords

Iron metabolism Laccase Lignification Phylogeny 

Abbreviations

BLAST

Basic local alignment search tool

CTAB

Cetyl trimethylammonium bromide

LMCO

Laccase-like multicopper oxidase

MCO

Multicopper oxidase

MPSS

Massively parallel signature sequencing

PCR

Polymerase chain reaction

pI

Isoelectric point

RACE

Rapid amplification of cDNA ends

RT-PCR

Reverse transcript polymerase chain reaction

TBE

Tris borate EDTA buffer

Notes

Acknowledgements

The authors wish to thank Chieh-Ting Wang for his assistance with tissue collection and RT-PCR assays. This work was supported by and NSF/Alfred P. Sloan Postdoctoral Fellowship (DBI-9803949) to B.C.M and a grant from the U.S. Department of Energy, Energy Biosciences Program (DE-FG02-99ER20336) to J.F.D.D.

References

  1. Appel RD, Bairoch A, Hochstrasser DF (1994) A new generation of information retrieval tools for biologists: the example of the ExPASy WWW server. Trends Biochem Sci 19:258–260CrossRefPubMedGoogle Scholar
  2. Aramayo R, Timberlake WE (1990) Sequence and molecular structure of the Aspergillus nidulans yA (laccase 1) gene. Nucleic Acids Res 18:3415–3415PubMedGoogle Scholar
  3. Bao W, O’Malley DM, Whetten R, Sederoff RR (1993) A laccase associated with lignification in loblolly pine xylem. Science 260:672–674Google Scholar
  4. Brouwers GJ, de Vrind JPM, Corstjens PLAM, Cornelis P, Baysse C, de Jong EWDV (1999) cumA, a gene encoding a multicopper oxidase, is involved in Mn2+ oxidation in Pseudomonas putida GB-1. Appl Environ Microb 65:1762–1768Google Scholar
  5. Butt VS (1980) Direct oxidases and related enzymes. In: Davies DD (ed) The biochemistry of plants, vol. 2. Academic, New York, pp 81–123Google Scholar
  6. Clegg MT, Cummings MP, Durbin ML (1997) The evolution of plant nuclear genes. Proc Natl Acad Sci USA 94:7791–7798CrossRefPubMedGoogle Scholar
  7. De Silva DM, Askwith CC, Eide D, Kaplan J (1995) The FET3 gene product required for high affinity iron transport in yeast is a cell surface ferroxidase. J Biol Chem 270:1098–1101CrossRefPubMedGoogle Scholar
  8. Dean JFD, Eriksson K-EL (1994) Laccase and the deposition of lignin in vascular plants. Holzforschung 48:21–33Google Scholar
  9. Dean JFD, LaFayette PR, Rugh C, Tristram AH, Hoopes JT, Merkle SA, Eriksson K-EL (1998) Laccases associated with lignifying tissues. In: Lewis NG, Sarkanen S (eds) Lignin and lignan biosynthesis. ACS Symposium Series 697:96–108Google Scholar
  10. Du YM, Oshima R, Kumanotani J (1984) Reversed-phase liquid-chromatographic separation and identification of constituents of urushiol in the sap of the lac tree, Rhus vernicifera. J Chromatogr 284:463–473CrossRefGoogle Scholar
  11. Eggert C, LaFayette PR, Temp U, Eriksson K-EL (1997) Laccase is essential for lignin degradation by the white-rot fungus Pycnoporus cinnabarinus. FEBS Lett 407:89–92CrossRefPubMedGoogle Scholar
  12. Emanuelsson O, Nielsen H, Brunak S, von Heijne G (2000) Predicting subcellular localization of proteins based on their N-terminal amino acid sequence. J Mol Biol 300:1005–1016PubMedGoogle Scholar
  13. Grass G, Rensing C (2001) Genes involved in copper homeostasis in Escherichia coli. J Bacteriol 183:2145–2147CrossRefPubMedGoogle Scholar
  14. Hebsgaard M, Korning PG, Tolstrup N, Engelbrecht J, Rouze P, Brunak S (1996) Splice site prediction in Arabidopsis thaliana DNA by combining local and global sequence information. Nucleic Acids Res 24:3439–3452CrossRefPubMedGoogle Scholar
  15. Hoopes JT, Dean JFD (2004) Ferroxidase activity in a laccase-like multicopper oxidase from Liriodendron tulipifera. Plant Physiol Biochem 42:27–33CrossRefPubMedGoogle Scholar
  16. Hüttermann A, Mai C, Kharazipour A (2001) Modification of lignin for the production of new compounded materials. Appl Microbiol Biotech 55:387–394CrossRefGoogle Scholar
  17. Kim C, Lorenz WW, Hoopes JT, Dean JFD (2001) Oxidation of phenolate siderophores by the multicopper oxidase encoded by the Escherichia coli yacK gene. J Bacteriol 183:4866–4875CrossRefPubMedGoogle Scholar
  18. LaFayette PR, Eriksson K-EL, Dean JFD (1999) Characterization and heterologous expression of laccase cDNAs from xylem tissues of yellow-poplar (Liriodendron tulipifera). Plant Mol Biol 40:23–35CrossRefPubMedGoogle Scholar
  19. Li W-H (1997) Molecular evolution. Sinauer Associates, Sunderland, pp 278–429Google Scholar
  20. Liu L, Dean JFD, Friedman WE, Eriksson K-EL (1994) A laccase-like phenoloxidase is correlated with lignin biosynthesis in Zinnia elegans stem tissues. Plant J 6:213–224CrossRefGoogle Scholar
  21. Martin W, Lydiate D, Brinkmann H, Forkmann G, Saedler H, Cerff R (1993) Molecular phylogenies in angiosperm evolution. Mol Biol Evol 10:140–162PubMedGoogle Scholar
  22. Mayer AM, Harel E (1979) Polyphenol oxidases in plants. Phytochemistry 18:193–215CrossRefGoogle Scholar
  23. Mayer AM, Staples RC (2002) Laccase: new functions for an old enzyme. Phytochemistry 60:551-565CrossRefPubMedGoogle Scholar
  24. Messerschmidt A (1997) Multi-copper oxidases. World Scientific, SingaporeGoogle Scholar
  25. Meyers BB, Lee DK, Vu TH, Tej SS, Edberg SB, Matvienko M, Tindell LD (2004) Arabidopsis MPSS: an online resource for quantitative expression analysis. Plant Physiol 135:801--813Google Scholar
  26. Musci G (2001) Ceruloplasmin, the unique multi-copper oxidase of vertebrates. Protein Peptide Lett 8:159–169CrossRefGoogle Scholar
  27. Nielsen H, Engelbrecht J, Brunak S, von Heijne G (1997) Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6Google Scholar
  28. O’Malley DM, Whetten R, Bao W, Chen CL, Sederoff RR (1993) The role of laccase in lignification. Plant J 4:751–757CrossRefGoogle Scholar
  29. Palmer AE, Randall DW, Xu F, Solomon EI (1999) Spectroscopic studies and electronic structure description of the high potential type 1 copper site in fungal laccase: Insight into the effect of the axial ligand. J Am Chem Soc 121:7138–7149CrossRefGoogle Scholar
  30. Parkin I, Sharpe A, Keith D, Lydiate D (1995) Identification of the A and C genomes of amphidiploid Brassica napus (oilseed rape). Genome 38:1122–1131Google Scholar
  31. Ranocha P, McDougall G, Hawkins S, Sterjiades R, Borderies G, Stewart D, Cabanes-Macheteau M, Boudet A-M, Goffner D (1999) Biochemical characterization, molecular cloning and expression of laccases—a divergent gene family—in poplar. Eur J Biochem 259:485–495CrossRefPubMedGoogle Scholar
  32. Salas SD, Bennett JE, Kwon-Chung KJ, Perfect JR, Williamson PR (1996) Effect of the laccase gene, CNLAC1, on virulence of Cryptococcus neoformans. J Exp Med 184:377–386CrossRefPubMedGoogle Scholar
  33. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning—a laboratory manual, 2nd edition. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  34. Sato Y, Bao WL, Sederoff R, Whetten R (2001) Molecular cloning and expression of eight laccase cDNAs in loblolly pine (Pinus taeda). J Plant Res 114:147–155Google Scholar
  35. Shi XL, Stoj C, Romeo A, Kosman DJ, Zhu ZW (2003) Fre1p Cu2+ reduction and fet3p Cu1+ oxidation modulate copper toxicity in Saccharomyces cerevisiae. J Biol Chem 278:50309–50315CrossRefPubMedGoogle Scholar
  36. Sterjiades R, Dean JFD, Eriksson K-EL (1992) Laccase from sycamore maple (Acer pseudoplatanus) polymerizes monolignols. Plant Physiol 99:1162–1168Google Scholar
  37. Stewart CN Jr, Via LE (1993) A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. BioTechniques 14:748–758PubMedGoogle Scholar
  38. Swofford DL (2000) PAUP*. Phylogenetic analysis using parsimony (*and other methods), version 4. Sinauer Associates, SunderlandGoogle Scholar
  39. Urbanowski JL, Piper RC (1999) The iron transporter fth1p forms a complex with the Fet5 iron oxidase and resides on the vacuolar membrane. J Biol Chem 274:38061–38070CrossRefPubMedGoogle Scholar
  40. vanWaasbergen LG, Hildebrand M, Tebo BM (1996) Identification and characterization of a gene cluster involved in manganese oxidation by spores of the marine Bacillus sp strain SG-1. J Bacteriol 178:3517–3530PubMedGoogle Scholar
  41. Xu F, Palmer AE, Yaver DS, Berka RM, Gambetta GA, Brown SH, Solomon EI (1999) Targeted mutations in a Trametes villosa laccase—axial perturbations of the T1 copper. J Biol Chem 274:12372–12375CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Bonnie C. McCaig
    • 1
    • 3
  • Richard B. Meagher
    • 2
  • Jeffrey F. D. Dean
    • 1
    Email author
  1. 1.Daniel B. Warnell School of Forest ResourcesUniversity of GeorgiaAthensUSA
  2. 2.Department of GeneticsUniversity of GeorgiaAthensUSA
  3. 3.Department of Energy-Plant Research LaboratoryMichigan State UniversityEast LansingUSA

Personalised recommendations