The small phytoplasma virulence effector SAP11 contains distinct domains required for nuclear targeting and CIN-TCP binding and destabilization

New Phytol. 2014 May;202(3):838-848. doi: 10.1111/nph.12721. Epub 2014 Feb 19.

Abstract

Phytoplasmas are insect-transmitted bacterial phytopathogens that secrete virulence effectors and induce changes in the architecture and defense response of their plant hosts. We previously demonstrated that the small (± 10 kDa) virulence effector SAP11 of Aster Yellows phytoplasma strain Witches' Broom (AY-WB) binds and destabilizes Arabidopsis CIN (CINCINNATA) TCP (TEOSINTE-BRANCHED, CYCLOIDEA, PROLIFERATION FACTOR 1 AND 2) transcription factors, resulting in dramatic changes in leaf morphogenesis and increased susceptibility to phytoplasma insect vectors. SAP11 contains a bipartite nuclear localization signal (NLS) that targets this effector to plant cell nuclei. To further understand how SAP11 functions, we assessed the involvement of SAP11 regions in TCP binding and destabilization using a series of mutants. SAP11 mutants lacking the entire N-terminal domain, including the NLS, interacted with TCPs but did not destabilize them. SAP11 mutants lacking the C-terminal domain were impaired in both binding and destabilization of TCPs. These SAP11 mutants did not alter leaf morphogenesis. A SAP11 mutant that did not accumulate in plant nuclei (SAP11ΔNLS-NES) was able to bind and destabilize TCP transcription factors, but instigated weaker changes in leaf morphogenesis than wild-type SAP11. Overall the results suggest that phytoplasma effector SAP11 has a modular organization in which at least three domains are required for efficient CIN-TCP destabilization in plants.

Keywords: TCP; coiled coil; effector; host target; mollicutes; phytoplasma; plant development; virulence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Arabidopsis / microbiology
  • Arabidopsis Proteins / metabolism*
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / metabolism*
  • Carrier Proteins / metabolism*
  • Cell Nucleus / metabolism*
  • Molecular Sequence Data
  • Mutation / genetics
  • Nuclear Export Signals
  • Phenotype
  • Phytoplasma / pathogenicity*
  • Plant Leaves / microbiology
  • Plant Leaves / physiology
  • Protein Binding
  • Protein Sorting Signals*
  • Protein Stability
  • Protein Structure, Tertiary
  • Saccharomyces cerevisiae / metabolism
  • Sequence Alignment
  • Structure-Activity Relationship
  • Virulence

Substances

  • Arabidopsis Proteins
  • Bacterial Proteins
  • Carrier Proteins
  • Nuclear Export Signals
  • Protein Sorting Signals