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Gene annotation: Metal transport & homeostasis Damien BLAUDEZ Annick BRUN Michel CHALOT

Gene annotation: Metal transport & homeostasis Damien BLAUDEZ Annick BRUN Michel CHALOT Barbara MONTANINI. UMR 1136 Interactions Arbres-Microorganismes. Metal transport & homeostasis in ECM fungi Why ?. At low metal concentrations: Tree nutrition is improved

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Gene annotation: Metal transport & homeostasis Damien BLAUDEZ Annick BRUN Michel CHALOT

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  1. Gene annotation: Metal transport & homeostasis Damien BLAUDEZ Annick BRUN Michel CHALOT Barbara MONTANINI UMR 1136 Interactions Arbres-Microorganismes

  2. Metal transport & homeostasis in ECM fungi Why ? At low metal concentrations: Tree nutrition is improved - higher metal import for trees (Cu, Mn, Zn, …) At toxic concentrations: Metal tolerance is improved - efficient sequestration in fungi - reduced export to trees (Cd, Pb, Zn, …)

  3. homeostasis tolerance Metals Metals

  4. Gene annotation strategy: Yeast as a model... by default

  5. Managing Zn homeostasis & tolerance...

  6. Managing Zn homeostasis & tolerance... Low affinity High affinity ZIP family

  7. high affinity Fungal ZIPs Lb = 5 gene models (+2 pseudogenes) Low affinity Vacuole ? Plasmalemma ?

  8. Zn transporter & Zn-finger transcription factor...

  9. Zn transporter & Zn-finger transcription factor...

  10. Managing Zn homeostasis & tolerance... CDF family (Cation diffusion facilitators)

  11. Fe Fungal CDFs Lb = 8 gene models Zn ?

  12. WT Pmr1D + PiCDF1 Pmr1D PiCDF1 : a key role in Mn tolerance ? Functional characterization in yeast Control + Mn Need further heterologous expression of Laccaria members

  13. Fe Fungal CDFs Zn Mn?

  14. Nramp family Yeast : 3 members Others B/A : 1 or 2 Laccaria : 1 vac Smf3p BLAST results: estExt_GeneWisePlus _worm.C_10257 = Smf1 ? (e-74) = Smf2 ? (e-73) = Smf3 ? (e-84) Other pathways for Mn uptake & homeostasis in Laccaria ? Role of Mn-CDFs ??? Managing Mn homeostasis & tolerance...

  15. Metallothioneins: 2 or 3 putative gene models (not predicted) Complexation Cd vacuole chélation (PC) PC occurence: - in plants - in S. pombe HMT1-like (ABC transporter) Managing Cd tolerance… ?

  16. Managing Cu homeostasis & tolerance...

  17. Is this fungus different ??

  18. Preliminary overview ... (Same picture with P-ATPase, ABC, MFS, MgTE,…)

  19. Preliminary overview ...

  20. Conclusions - Presence of the major pathways... - No phytochelatin synthesis pathway ! - New sub-group in the CDF family (Mn transport ?) - Gene numbers: no major difference with other filamentous fungi - Alternative splicing ? Too few ESTs !! - Gene models not predicted for small proteins (metallothioneins)

  21. Mn-CDFs: Where do they work ? Managing Mn tolerance... Smf1/2

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