GAP45 phosphorylation controls assembly of the Toxoplasma myosin XIV complex

Stacey D. Gilk, Elizabeth Gaskins, Gary E. Ward, Con J.M. Beckers

Research output: Contribution to journalArticlepeer-review

57 Scopus citations

Abstract

Toxoplasma gondii motility is powered by the myosin XIV motor complex, which consists of the myosin XIV heavy chain (MyoA), the myosin light chain (MLC1), GAP45, and GAP50, the membrane anchor of the complex. MyoA, MLC1, and GAP45 are initially assembled into a soluble complex, which then associates with GAP50, an integral membrane protein of the parasite inner membrane complex. While all proteins in the myosin XIV motor complex are essential for parasite survival, the specific role of GAP45 remains unclear. We demonstrate here that final assembly of the motor complex is controlled by phosphorylation of GAP45. This protein is phosphorylated on multiple residues, and by using mass spectroscopy, we have identified two of these, Ser163 and Ser 167. The importance of these phosphorylation events was determined by mutation of Ser163 and Ser167 to Glu and Ala residues to mimic phosphorylated and nonphosphorylated residues, respectively. Mutation of Ser163 and Ser167 to either Ala or Glu residues does not affect targeting of GAP45 to the inner membrane complex or its association with MyoA and MLC1. Mutation of Ser163 and Ser167 to Ala residues also does not affect assembly of the mutant GAP45 protein into the myosin motor complex. Mutation of Ser163 and Ser167 to Glu residues, however, prevents association of the MyoA-MLC1-GAP45 complex with GAP50. These observations indicate that phosphorylation of Ser163 and Ser167 in GAP45 controls the final step in assembly of the myosin XIV motor complex.

Original languageEnglish (US)
Pages (from-to)190-196
Number of pages7
JournalEukaryotic Cell
Volume8
Issue number2
DOIs
StatePublished - Feb 2009
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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