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Gene annotation: Laccases and multicopper oxidases

Gene annotation: Laccases and multicopper oxidases. 4th Workshop of the Laccaria Genome Consortium Georg-August-Universität, Göttingen - November 16-17, 2006. P.J. Hoegger 1 , S. Kilaru 1 , P.E. Courty 2 , F. Martin 2 , U. Kües 1 1 Georg-August-University Göttingen, Germany

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Gene annotation: Laccases and multicopper oxidases

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  1. Gene annotation: Laccases and multicopper oxidases 4th Workshop of the Laccaria Genome Consortium Georg-August-Universität, Göttingen - November 16-17, 2006 P.J. Hoegger1, S. Kilaru1, P.E. Courty2, F. Martin2, U. Kües1 1Georg-August-University Göttingen, Germany 2INRA Nancy, France

  2. Multi-copper oxidase family main members: - laccases - fungi - plants - insects - bacteria - ascorbate oxidases - plants - ferroxidases - fungi - ceruloplasmins - mammals

  3. Laccase function and application Biological functions: - lignin degradation - lignification - pigment synthesis - fruiting body formation - melanin polymerization - detoxification - cuticle sclerotization - soil litter polymer degrad. Applications: - bleaching processes - bioremediation - composite wood production

  4. 1 type-2 Cu 2 type-3 Cu 1 type-1 Cu Trametes versicolor laccase Lcc1 Piontek et al. 2002, JBC 277, 37663-37669

  5. Substrate binding loops I III II IV Fungal laccase structure Signal sequence H H H H H H H HCH H L1 L2 L3 L4 signature sequences, including copper ligands

  6. lcc11 lcc1 222 kb 262 kb 25 kb lcc2 73 kb 175 kb lcc3 112 kb 3166 kb lcc4 194 kb 904 kb lcc5 116 kb 312 kb lcc9 143 kb 110 kb lcc10 647 kb 65 kb lcc6 lcc7 lcc8 429 kb 569 kb 0.53 kb 0.53 kb MCOs in the genome of L. bicolor Scaffold 39 Scaffold 2 Scaffold 69 Scaffold 47 Scaffold 13 Scaffold 14 Scaffold 67 Scaffold 27

  7. basidiomycete laccases laccases sensu stricto ascomycete laccases insect laccases Cel NP 501502 fungal pigment MCOs fungal ferroxidases Mgr Mco7 Sce AAB64948 Cgl XP 448078 Kla XP 452271 ascorbate oxidases plant LMCOs Pch AAO42609 MCO1 Pch AAS21669 MCO4 Pch AAS21659 MCO2 Pch AAS21662 MCO3 Mgr Mco1 Fgr Mco1 "ferroxidases / laccases" Uma Mco1 Ego NP 984335 Uma Mco3 Cne Mco5 Cne A36962 Cne Mco6 Cim Mco2 Fgr Mco10 bacterial MCOs 0.1 Multi-copper oxidase phylogenetic tree

  8. LbiLcc1 LbiLcc2 LbiLcc4 LbiLcc3 basidiomycete laccases laccases sensu stricto LbiLcc6 LbiLcc5 LbiLcc7 LbiLcc8 LbiLcc9 ascomycete laccases insect laccases Cel NP 501502 fungal pigment MCOs fungal ferroxidases LbiLcc10 Mgr Mco7 LbiLcc11 Sce AAB64948 Cgl XP 448078 Kla XP 452271 ascorbate oxidases plant LMCOs Pch AAO42609 MCO1 Pch AAS21669 MCO4 Pch AAS21659 MCO2 Pch AAS21662 MCO3 Mgr Mco1 Fgr Mco1 "ferroxidases / laccases" Uma Mco1 Ego NP 984335 Uma Mco3 Cne Mco5 Cne A36962 Cne Mco6 Cim Mco2 Fgr Mco10 bacterial MCOs 0.1 L. bicolor MCOs in phylogenetic tree

  9. wood decaying species (Aphyllophorales, Agaricales) wood and litter decomposers (Agaricales)

  10. Intron position conservation in basidiomycete laccase genes

  11. Tree based on intron positions

  12. lcc11 lcc1 222 kb 262 kb 25 kb lcc2 73 kb 175 kb lcc3 112 kb 3166 kb lcc4 194 kb 904 kb lcc5 116 kb 312 kb lcc9 143 kb 110 kb lcc10 647 kb 65 kb lcc6 lcc7 lcc8 429 kb 569 kb 0.53 kb 0.53 kb L. bicolor MCOgene family Scaffold 39 Scaffold 2 Scaffold 69 Scaffold 47 Scaffold 13 Scaffold 14 Scaffold 67 Scaffold 27

  13. Syn. / non-syn. substitution rates

  14. LbiLcc1 LbiLcc2 LbiLcc4 LbiLcc3 basidiomycete laccases laccases sensu stricto LbiLcc6 LbiLcc5 LbiLcc7 LbiLcc8 LbiLcc9 ascomycete laccases insect laccases Cel NP 501502 fungal pigment MCOs fungal ferroxidases LbiLcc10 Mgr Mco7 LbiLcc11 Sce AAB64948 Cgl XP 448078 Kla XP 452271 ascorbate oxidases plant LMCOs Pch AAO42609 MCO1 Pch AAS21669 MCO4 Pch AAS21659 MCO2 Pch AAS21662 MCO3 Mgr Mco1 Fgr Mco1 "ferroxidases / laccases" Uma Mco1 Ego NP 984335 Uma Mco3 Cne Mco5 Cne A36962 Cne Mco6 Cim Mco2 Fgr Mco10 bacterial MCOs 0.1 L. bicolor MCOs in phylogenetic tree

  15. Iron uptake in fungi Kosman 2003, Mol Microbiol, 47:1185-1197

  16. lcc10 and lcc11 loci iron permease ferroxidase 179 bp 331 bp 115 bp ftr1a lcc10 nucleotide id. 89% 86% aa id. / sim. 91% / 95% 84% / 88% ftr1b lcc11 205 bp

  17. Syn. / non-syn. substitution rates of lcc10/lcc11 and ftr1a/ftr1b

  18. Intron position conservation in basidiomycete ferroxidase genes

  19. MCO arsenal inbasidiomycete species

  20. MCO arsenal inbasidiomycete species

  21. MCO expression profile

  22. Acknowledgements Molecular Wood Biotechnology, Institute of Forest Botany, Georg-August-University Göttingen, Germany Ursula Kües Patrik Hoegger Sreedhar Kilaru UMR 1136 Interactions Arbres Micro-organismes, Centre INRA de Nancy, Champenoux, France Francis Martin Marc Buée Pierre-Emmanuel Courty

  23. MCO arsenal in fungal species basidiomycetes filamentous ascomycetes ascomycetous yeasts z

  24. A/C>T A>G frame- shift? T/C>G A>G G>A lcc10 and lcc11 loci 179 bp 331 bp 115 bp ftr1a lcc10 nucleotide id. 89% 86% aa id. / sim. 91% / 95% 84% / 88% ftr1b lcc11 205 bp

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