PXD039432

PXD039432 is an original dataset announced via ProteomeXchange.

Title	Analysis of CDPK1 targets identifies a trafficking adaptor complex that regulates microneme exocytosis in Toxoplasma - HOOK and FTS IPs
Description	Apicomplexan parasites use Ca2+-regulated exocytosis to secrete essential virulence factors from specialized organelles called micronemes. Ca2+-dependent protein kinases (CDPKs) are required for microneme exocytosis; however, the molecular events that regulate trafficking and fusion of micronemes with the plasma membrane remain unresolved. Here, we combine sub-minute resolution phosphoproteomics and bio-orthogonal labeling of kinase substrates in Toxoplasma gondii to identify 163 proteins phosphorylated in a CDPK1-dependent manner. In addition to known regulators of secretion, we identify uncharacterized targets with predicted functions across signaling, gene expression, trafficking, metabolism, and ion homeostasis. One of the CDPK1 targets is a putative HOOK activating adaptor. In other eukaryotes, HOOK homologs form the FHF complex with FTS and FHIP to activate dynein-mediated trafficking of endosomes along microtubules. We show the FHF complex is partially conserved in T. gondii, consisting of HOOK, an FTS homolog, and two parasite-specific proteins (TGGT1_306920 and TGGT1_316650). CDPK1 kinase activity and HOOK are required for the rapid apical trafficking of micronemes as parasites initiate motility. Moreover, parasites lacking HOOK or FTS display impaired microneme protein secretion, leading to a block in the invasion of host cells. Taken together, our work provides a comprehensive catalog of CDPK1 targets and reveals how vesicular trafficking has been tuned to support a parasitic lifestyle.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:26:27.597.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD039432
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alex Chan
SpeciesList	scientific name: Toxoplasma gondii GT1; NCBI TaxID: 507601;
ModificationList	methylthiolated residue; phosphorylated residue; acetylated residue; monohydroxylated residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-01-13 14:25:57	ID requested
1	2023-10-18 10:51:41	announced
⏵ 2	2023-11-14 08:26:27	announced	2023-11-14: Updated project metadata.

10.6019/PXD039432;

submitter keyword: T. gondii, LC-MSMS, vesicular trafficking, signaling

Dr. Sebastian Lourido
contact affiliation	Associate Professor, Department of Biology, Massachusetts Institute of Technology and Whitehead Institute of Biomedical Research
contact email	lourido@wi.mit.edu
lab head
Alex Chan
contact affiliation	Whitehead Institute for Biomedical Research Department of Biology, Massachusetts Insitute of Technology
contact email	awchan@wi.mit.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD039432
     1. Label: PRIDE project
     2. Name: Analysis of CDPK1 targets identifies a trafficking adaptor complex that regulates microneme exocytosis in Toxoplasma - HOOK and FTS IPs