Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation.
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 (...
Guardado en:
| Autores principales: | , , , , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Public Library of Science (PLoS)
2014
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/f1ae95746a6f44ef882a37a662a2b130 |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:f1ae95746a6f44ef882a37a662a2b130 |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:f1ae95746a6f44ef882a37a662a2b1302021-11-18T06:07:09ZLysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation.1553-73661553-737410.1371/journal.ppat.1003830https://doaj.org/article/f1ae95746a6f44ef882a37a662a2b1302014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24391497/?tool=EBIhttps://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Histone 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.Jiachen WangStacy E DixonLi-Min TingTing-Kai LiuVictoria JeffersMatthew M CrokenMyrasol CallowayDominique CannellaMohamed Ali HakimiKami KimWilliam J SullivanPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 10, Iss 1, p e1003830 (2014) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 |
| spellingShingle |
Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 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 Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. |
| description |
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. |
| format |
article |
| author |
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 |
| author_facet |
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 |
| author_sort |
Jiachen Wang |
| title |
Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. |
| title_short |
Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. |
| title_full |
Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. |
| title_fullStr |
Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. |
| title_full_unstemmed |
Lysine acetyltransferase GCN5b interacts with AP2 factors and is required for Toxoplasma gondii proliferation. |
| title_sort |
lysine acetyltransferase gcn5b interacts with ap2 factors and is required for toxoplasma gondii proliferation. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2014 |
| url |
https://doaj.org/article/f1ae95746a6f44ef882a37a662a2b130 |
| work_keys_str_mv |
AT jiachenwang lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT stacyedixon lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT liminting lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT tingkailiu lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT victoriajeffers lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT matthewmcroken lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT myrasolcalloway lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT dominiquecannella lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT mohamedalihakimi lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT kamikim lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation AT williamjsullivan lysineacetyltransferasegcn5binteractswithap2factorsandisrequiredfortoxoplasmagondiiproliferation |
| _version_ |
1718424577338507264 |