A Quantitative, High-Throughput Reverse Genetic Screen Reveals Novel Connections Between pre-mRNA Splicing and 5’ and 3’ end Transcript Determinants

ABSTRACT: The coding portions of most eukaryotic genes are interrupted by non-coding regions termed introns that must be excised prior to their translation. The excision of introns from precursor messenger RNA (pre-mRNA), is catalyzed by the spliceosome, a large macromolecule composed of both RNA and protein components. Several studies have uncovered connections between pre-mRNA splicing and other RNA processing pathways such as the remodeling of chromatin structure, transcription and processing events which take place at the 3’ end of the transcript. To date, however, the full complement of factors which function to couple splicing to other processes in the cell remains unknown. Here, we have developed a novel screening methodology in the budding yeast, Saccharomyces cerevisiae, that allowed us to individually examine nearly all of the ~6000 genes to determine which factors functionally impact splicing. We identified mutations in components that function at either the 5' or 3' end of a gene. Most of these components have previously established roles in other aspects of gene expression, including chromatin remodeling and cleavage and polyadenylation processes, and their identification here provides the first evidence for their roles in coupling these pathways. Included here are both the QPCR data generated from this high-throughput reverse genetic screen, as well as the microarray analyses of several of the candidate factors identified by the screen.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Jeffrey Pleiss 

PROVIDER: E-GEOD-34330 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress