title: Gene expression regulatory networks in Trypanosoma brucei: insights into the role of the mRNA-binding proteome.
abstract: Control of gene expression at the post-transcriptional level is essential in all organisms, and RNA-binding proteins play critical roles from mRNA synthesis to decay. To fully understand this process, it is necessary to identify the complete set of RNA-binding proteins and the functional consequences of the protein-mRNA interactions. Here, we provide an overview of the proteins that bind to mRNAs and their functions in the pathogenic bloodstream form of Trypanosoma brucei. We describe the production of a small collection of open-reading frames encoding proteins potentially involved in mRNA metabolism. With this ORFeome collection, we used tethering to screen for proteins that play a role in post-transcriptional control. A yeast two-hybrid screen showed that several of the discovered repressors interact with components of the CAF1/NOT1 deadenylation complex. To identify the RNA-binding proteins, we obtained the mRNA-bound proteome. We identified 155 high-confidence candidates, including many not previously annotated as RNA-binding proteins. Twenty seven of these proteins affected reporter expression in the tethering screen. Our study provides novel insights into the potential trypanosome mRNPs composition, architecture and function.
authors.affiliation: Functional Genome Analysis, Deutsche Krebsforschungszentrum (DKFZ), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany.
authors.forename: Smiths
authors.initials: S
authors.lastname: Lueong
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282.
authors.forename: Clementine
authors.initials: C
authors.lastname: Merce
authors.affiliation: Computational Genome Biology, Deutsches Krebsforschungszentrum (DKFZ), Im Neuenheimer Feld 580, 69120 Heidelberg.
authors.forename: Bernd
authors.initials: B
authors.lastname: Fischer
authors.affiliation: Functional Genome Analysis, Deutsche Krebsforschungszentrum (DKFZ), Im Neuenheimer Feld 580, 69120 Heidelberg, Germany.
authors.forename: Jörg D
authors.initials: JD
authors.lastname: Hoheisel
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282.
authors.forename: Esteban D
authors.initials: ED
authors.lastname: Erben
mesh_terms: D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D057166:High-Throughput Screening Assays; D020543:Proteome / Q000235:genetics*; D015800:Protozoan Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics*; D016053:RNA, Protozoan / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000235:genetics*
chemical_list: D020543:Proteome; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D016601:RNA-Binding Proteins