DOI: 10.1128/ec.00201-08. GAP45 Phosphorylation Controls Assembly of the <i>Toxoplasma</i> Myosin XIV Complex

GAP45 Phosphorylation Controls Assembly of the Toxoplasma Myosin XIV Complex

10.1128/ec.00201-08

Crossref journal-article

American Society for Microbiology

Eukaryotic Cell (235)

Abstract

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, Ser 163 and Ser 167 . The importance of these phosphorylation events was determined by mutation of Ser 163 and Ser 167 to Glu and Ala residues to mimic phosphorylated and nonphosphorylated residues, respectively. Mutation of Ser 163 and Ser 167 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 Ser 163 and Ser 167 to Ala residues also does not affect assembly of the mutant GAP45 protein into the myosin motor complex. Mutation of Ser 163 and Ser 167 to Glu residues, however, prevents association of the MyoA-MLC1-GAP45 complex with GAP50. These observations indicate that phosphorylation of Ser 163 and Ser 167 in GAP45 controls the final step in assembly of the myosin XIV motor complex.

Bibliography

Gilk, S. D., Gaskins, E., Ward, G. E., & Beckers, C. J. M. (2009). GAP45 Phosphorylation Controls Assembly of the Toxoplasma Myosin XIV Complex. Eukaryotic Cell, 8(2), 190â196.

Authors 4

Stacey D. Gilk (first)
Elizabeth Gaskins (additional)
Gary E. Ward (additional)
Con J. M. Beckers (additional)

References 27 Referenced 67

Dates

Type	When
Created	16 years, 8 months ago (Dec. 1, 2008, 8:54 p.m.)
Deposited	3 years, 6 months ago (Feb. 23, 2022, 9:41 a.m.)
Indexed	4 weeks, 2 days ago (Aug. 2, 2025, 12:36 a.m.)
Issued	16 years, 7 months ago (Feb. 1, 2009)
Published	16 years, 7 months ago (Feb. 1, 2009)
Published Print	16 years, 7 months ago (Feb. 1, 2009)

Funders 0

None

BibTeX

@article{Gilk_2009, title={GAP45 Phosphorylation Controls Assembly of the Toxoplasma Myosin XIV Complex}, volume={8}, ISSN={1535-9786}, url={http://dx.doi.org/10.1128/ec.00201-08}, DOI={10.1128/ec.00201-08}, number={2}, journal={Eukaryotic Cell}, publisher={American Society for Microbiology}, author={Gilk, Stacey D. and Gaskins, Elizabeth and Ward, Gary E. and Beckers, Con J. M.}, year={2009}, month=feb, pages={190–196} }

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  "abstract": "<jats:title>ABSTRACT</jats:title>\n          <jats:p>\n            <jats:italic>Toxoplasma gondii</jats:italic>\n            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, Ser\n            <jats:sup>163</jats:sup>\n            and Ser\n            <jats:sup>167</jats:sup>\n            . The importance of these phosphorylation events was determined by mutation of Ser\n            <jats:sup>163</jats:sup>\n            and Ser\n            <jats:sup>167</jats:sup>\n            to Glu and Ala residues to mimic phosphorylated and nonphosphorylated residues, respectively. Mutation of Ser\n            <jats:sup>163</jats:sup>\n            and Ser\n            <jats:sup>167</jats:sup>\n            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 Ser\n            <jats:sup>163</jats:sup>\n            and Ser\n            <jats:sup>167</jats:sup>\n            to Ala residues also does not affect assembly of the mutant GAP45 protein into the myosin motor complex. Mutation of Ser\n            <jats:sup>163</jats:sup>\n            and Ser\n            <jats:sup>167</jats:sup>\n            to Glu residues, however, prevents association of the MyoA-MLC1-GAP45 complex with GAP50. These observations indicate that phosphorylation of Ser\n            <jats:sup>163</jats:sup>\n            and Ser\n            <jats:sup>167</jats:sup>\n            in GAP45 controls the final step in assembly of the myosin XIV motor complex.\n          </jats:p>",
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