GSE118232
Genome-wide function of MCM-BP in Trypanosoma brucei DNA replication and transcription
In Trypanosoma brucei, genes are arranged in Polycistronic Transcription Units (PTUs), which are demarcated by transcription start and stop sites. Transcription start sites are also binding sites of Origin Recognition Complex 1 (ORC1). These suggest that transcription and replication in trypanosomes must occur in highly ordered and cooperative manners. Not coincidently, our previous genetic screen, a LOss of transcription Silencing (LOS) screen, identified a T. brucei MCM-BP, which forms a complex with subunits of the replicative helicase MCM. Here, I show that TbMCM-BP is required for DNA replication and transcription. TbMCM-BP depletion causes a significant reduction of replicating cells in S phase and genome-wide impairments of replication origin activation. Moreover, levels of sense and antisense transcripts increase at boundaries of PTUs in the absence of TbMCM-BP. TbMCM-BP is also important for transcriptional repression of the specialized subtelomeric PTUs, the Bloodstream-form Expression-Sites (BESs), which house the gene of antigenic importance, VSG. In the absence of TbMCM-BP, expression of silent VSGs is elevated, silent promoter regions are derepressed, and antisense transcription increases downstream of the silent promoters.  This study reveals that TbMCM-BP, a replication initiation protein, guides transcription to properly start and stop, and also helps it move in the correct direction. Overall design: For RNAseq, sample for each time point have four replicates. For MFAseq, one sample per each time point was used, and FACS sorted at G1, early S (S1), late S (S2) and G2. Please note that the associated processed data files are indicated in the corresponding sample description field and the readme.txt contains a brief description of the data content for each file.