Structural basis for PI(4)P-specific membrane recruitment of the Legionella pneumophila effector DrrA/SidM

Structure. 2014 Mar 4;22(3):397-408. doi: 10.1016/j.str.2013.12.018. Epub 2014 Feb 13.

Abstract

Recruitment of the Legionella pneumophila effector DrrA to the Legionella-containing vacuole, where it activates and AMPylates Rab1, is mediated by a P4M domain that binds phosphatidylinositol 4-phosphate [PI(4)P] with high affinity and specificity. Despite the importance of PI(4)P in Golgi trafficking and its manipulation by pathogens, the structural bases for PI(4)P-dependent membrane recruitment remain poorly defined. Here, we determined the crystal structure of a DrrA fragment including the P4M domain in complex with dibutyl PI(4)P and investigated the determinants of phosphoinositide recognition and membrane targeting. Headgroup recognition involves an elaborate network of direct and water-mediated interactions with basic and polar residues in the context of a deep, constrictive binding pocket. An adjacent hydrophobic helical element packs against the acyl chains and inserts robustly into PI(4)P-containing monolayers. The structural, biochemical, and biophysical data reported here support a detailed structural mechanism for PI(4)P-dependent membrane targeting by DrrA.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Binding Sites
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • Crystallography, X-Ray
  • Guanine Nucleotide Exchange Factors / chemistry*
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Legionella pneumophila / chemistry
  • Legionella pneumophila / metabolism
  • Models, Molecular
  • Phosphatidylinositol Phosphates / metabolism*
  • Protein Conformation

Substances

  • Bacterial Proteins
  • Guanine Nucleotide Exchange Factors
  • Phosphatidylinositol Phosphates
  • SidM protein, Legionella pneumophila
  • phosphatidylinositol 4-phosphate

Associated data

  • PDB/4MXP