Functional Genomic Analysis of Novel Cell Cycle Transcription Factors in Schizosaccharomyce Pombe

Abstract

The transcriptional-regulatory network in Schizosaccharomyces pombe remains poorly characterized with approximately 10% of transcription factors being well characterized. Attempts to complete the mapping of the transcriptional-regulatory network have been hindered by the observation that almost 90% of transcription factor deletions do not exhibit growth defects in rich media. To circumvent this issue, we have overexpressed six HA-tagged transcription factors (SPAC25B8.19, SPCC1393.08, phx1+, toe2+, toe3+, and toe4+) under the control of the nmt41 promoter. In order to identify the direct targets of these transcription factors, we performed expression microarray profiling, chromatin immunoprecipitation chip, qPCR, phenotypic replication, and genetic rescue on these strains. Gene ontology searching and motif finding to identify functional enrichment and conserved regulatory motifs were also performed. Through these techniques, we were able to describe putative roles for involvement of five transcription factors in the regulation of retrotransposable elements, meiosis, long term survival, septation, nuclear segregation, and cellular proliferation.