The bromodomain-containing protein Ibd1 links multiple chromatin-related protein complexes to highly expressed genes in Tetrahymena thermophila.
BACKGROUND
The chromatin remodelers of the SWI/SNF family are critical transcriptional regulators. Recognition of lysine acetylation through a bromodomain (BRD) component is key to SWI/SNF function; in most eukaryotes, this function is attributed to SNF2/Brg1.


RESULTS
Using affinity purification coupled to mass spectrometry (AP-MS) we identified members of a SWI/SNF complex (SWI/SNFTt) in Tetrahymena thermophila. SWI/SNFTt is composed of 11 proteins, Snf5Tt, Swi1Tt, Swi3Tt, Snf12Tt, Brg1Tt, two proteins with potential chromatin-interacting domains and four proteins without orthologs to SWI/SNF proteins in yeast or mammals. SWI/SNFTt subunits localize exclusively to the transcriptionally active macronucleus during growth and development, consistent with a role in transcription. While Tetrahymena Brg1 does not contain a BRD, our AP-MS results identified a BRD-containing SWI/SNFTt component, Ibd1 that associates with SWI/SNFTt during growth but not development. AP-MS analysis of epitope-tagged Ibd1 revealed it to be a subunit of several additional protein complexes, including putative SWRTt, and SAGATt complexes as well as a putative H3K4-specific histone methyl transferase complex. Recombinant Ibd1 recognizes acetyl-lysine marks on histones correlated with active transcription. Consistent with our AP-MS and histone array data suggesting a role in regulation of gene expression, ChIP-Seq analysis of Ibd1 indicated that it primarily binds near promoters and within gene bodies of highly expressed genes during growth.


CONCLUSIONS
Our results suggest that through recognizing specific histones marks, Ibd1 targets active chromatin regions of highly expressed genes in Tetrahymena where it subsequently might coordinate the recruitment of several chromatin-remodeling complexes to regulate the transcriptional landscape of vegetatively growing Tetrahymena cells.
['Saettone|Alejandro|A|', 'Garg|Jyoti|J|', 'Lambert|Jean-Philippe|JP|', 'Nabeel-Shah|Syed|S|', 'Ponce|Marcelo|M|', 'Burtch|Alyson|A|', 'Thuppu Mudalige|Cristina|C|', 'Gingras|Anne-Claude|AC|', 'Pearlman|Ronald E|RE|', 'Fillingham|Jeffrey|J|0000-0002-9000-1458']
D044127:Epigenesis, Genetic D005786:Gene Expression Regulation D018507:Gene Expression Regulation, Developmental D006657:Histones D013058:Mass Spectrometry D009687:Nuclear Proteins D040901:Proteomics D015800:Protozoan Proteins D016808:Tetrahymena thermophila D014158:Transcription, Genetic
Tetrahymena Bromodomain Chromatin-remodeling complexes