Lysine Acetyltransferase GCN5b Interacts with AP2 Factors and Is Required for Toxoplasma gondii Proliferation

Jiachen Wang, Stacy E. Dixon, Li Min Ting, Ting Kai Liu, Victoria Jeffers, Matthew M. Croken, Myrasol Calloway, Dominique Cannella, Mohamed Ali Hakimi, Kami Kim, William J. Sullivan

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Histone acetylation has been linked to developmental changes in gene expression and is a validated drug target of apicomplexan parasites, but little is known about the roles of individual histone modifying enzymes and how they are recruited to target genes. The protozoan parasite Toxoplasma gondii (phylum Apicomplexa) is unusual among invertebrates in possessing two GCN5-family lysine acetyltransferases (KATs). While GCN5a is required for gene expression in response to alkaline stress, this KAT is dispensable for parasite proliferation in normal culture conditions. In contrast, GCN5b cannot be disrupted, suggesting it is essential for Toxoplasma viability. To further explore the function of GCN5b, we generated clonal parasites expressing an inducible HA-tagged dominant-negative form of GCN5b containing a point mutation that ablates enzymatic activity (E703G). Stabilization of this dominant-negative GCN5b was mediated through ligand-binding to a destabilization domain (dd) fused to the protein. Induced accumulation of the ddHAGCN5b(E703G) protein led to a rapid arrest in parasite replication. Growth arrest was accompanied by a decrease in histone H3 acetylation at specific lysine residues as well as reduced expression of GCN5b target genes in GCN5b(E703G) parasites, which were identified using chromatin immunoprecipitation coupled with microarray hybridization (ChIP-chip). Proteomics studies revealed that GCN5b interacts with AP2-domain proteins, apicomplexan plant-like transcription factors, as well as a "core complex" that includes the co-activator ADA2-A, TFIID subunits, LEO1 polymerase-associated factor (Paf1) subunit, and RRM proteins. The dominant-negative phenotype of ddHAGCN5b(E703G) parasites, considered with the proteomics and ChIP-chip data, indicate that GCN5b plays a central role in transcriptional and chromatin remodeling complexes. We conclude that GCN5b has a non-redundant and indispensable role in regulating gene expression required during the Toxoplasma lytic cycle.

Original languageEnglish (US)
Article numbere1003830
JournalPLoS Pathogens
Volume10
Issue number1
DOIs
StatePublished - Jan 2014

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Toxoplasma
Parasites
Histones
Acetylation
Gene Expression
Proteomics
Transcription Factor TFIID
Apicomplexa
Plant Proteins
Chromatin Assembly and Disassembly
Chromatin Immunoprecipitation
Invertebrates
Lysine Acetyltransferases
Point Mutation
Genes
Lysine
Proteins
Transcription Factors
Ligands
Phenotype

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

Wang, J., Dixon, S. E., Ting, L. M., Liu, T. K., Jeffers, V., Croken, M. M., ... Sullivan, W. J. (2014). Lysine Acetyltransferase GCN5b Interacts with AP2 Factors and Is Required for Toxoplasma gondii Proliferation. PLoS Pathogens, 10(1), [e1003830]. https://doi.org/10.1371/journal.ppat.1003830

Lysine Acetyltransferase GCN5b Interacts with AP2 Factors and Is Required for Toxoplasma gondii Proliferation. / Wang, Jiachen; Dixon, Stacy E.; Ting, Li Min; Liu, Ting Kai; Jeffers, Victoria; Croken, Matthew M.; Calloway, Myrasol; Cannella, Dominique; Ali Hakimi, Mohamed; Kim, Kami; Sullivan, William J.

In: PLoS Pathogens, Vol. 10, No. 1, e1003830, 01.2014.

Research output: Contribution to journalArticle

Wang, J, Dixon, SE, Ting, LM, Liu, TK, Jeffers, V, Croken, MM, Calloway, M, Cannella, D, Ali Hakimi, M, Kim, K & Sullivan, WJ 2014, 'Lysine Acetyltransferase GCN5b Interacts with AP2 Factors and Is Required for Toxoplasma gondii Proliferation', PLoS Pathogens, vol. 10, no. 1, e1003830. https://doi.org/10.1371/journal.ppat.1003830
Wang, Jiachen ; Dixon, Stacy E. ; Ting, Li Min ; Liu, Ting Kai ; Jeffers, Victoria ; Croken, Matthew M. ; Calloway, Myrasol ; Cannella, Dominique ; Ali Hakimi, Mohamed ; Kim, Kami ; Sullivan, William J. / Lysine Acetyltransferase GCN5b Interacts with AP2 Factors and Is Required for Toxoplasma gondii Proliferation. In: PLoS Pathogens. 2014 ; Vol. 10, No. 1.
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AU - Jeffers, Victoria

AU - Croken, Matthew M.

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