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Role of the ELL complex in transcriptional regulation in S. pombe
Author(s) -
Gopalan Sneha,
Gibbon Dana,
Seidel Chris,
Zhang Ying,
Florens Laurence,
Washburn Michael,
Conaway Joan,
Conaway Ronald
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.593.12
Subject(s) - gene , schizosaccharomyces pombe , transcription (linguistics) , rna polymerase ii , genetics , transcription factor , biology , transcription factor ii e , promoter , transcriptional regulation , saccharomyces cerevisiae , microbiology and biotechnology , gene expression , linguistics , philosophy
The ELL family of transcription elongation factors can suppress the transient pausing of Pol II at each step of nucleotide addition during transcription, thereby stimulating the overall rate of transcription. ELL‐associated factors (EAFs) bind directly to ELL and are positive regulators of ELL elongation activity. ELL and EAF have been implicated in the release of RNA pol II from promoter proximal pausing by an unknown mechanism. Orthologs of ELL and EAF were not identified in fungi until recently. Previous data from our lab identified genes in S. pombe similar to ELL and EAF, which have no obvious orthologs in S. cerevisiae . SpELL and SpEAF also interact with each other to form a heterodimer and can positively regulate transcription by RNA pol II. We identified proteins interacting with SpELL and SpEAF by immunoprecipitation followed by Multidimensional Protein Identification Technology (MudPIT). Proteomic analyses revealed an uncharacterized sequence orphan protein that associates strongly with both ELL and EAF. Biochemical analysis using recombinant proteins confirmed that this protein binds directly to ELL and forms a ternary complex with ELL and EAF. ChIP‐seq experiments suggest that all three proteins co‐localize at genes with high RNA pol II occupancy. Synthetic Genetic Array (SGA), a high‐throughput technique for exploring genetic interactions, was performed on the deletion mutants lacking the genes encoding ELL, EAF and the novel ELL‐interacting protein. Preliminary data analysis suggests that these genes may interact genetically with overlapping set of genes. Support or Funding Information