High throughput sequencing analysis of Trypanosoma brucei DRBD3/PTB1-bound mRNAs.
Trypanosomes are early-branched eukaryotes that show an unusual dependence on post-transcriptional mechanisms to regulate gene expression. RNA-binding proteins are crucial in controlling mRNA fate in these organisms, but their RNA substrates remain largely unknown. Here we have analyzed on a global scale the mRNAs associated with the Trypanosoma brucei RNA-binding protein DRBD3/PTB1, by capturing ribonucleoprotein complexes using UV cross-linking and subsequent immunoprecipitation. DRBD3/PTB1 associates with many transcripts encoding ribosomal proteins and translation factors. Consequently, silencing of DRBD3/PTB1 expression altered the protein synthesis rate. DRBD3/PTB1 also binds to mRNAs encoding the enzymes required to obtain energy through the oxidation of proline to succinate. We hypothesize that DRBD3/PTB1 is a key player in RNA regulon-based gene control influencing protein synthesis in trypanosomes.
Department of Microbiology and Molecular Genetics, Rutgers-New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ, USA.
Anish
A
Das
Department of Microbiology and Molecular Genetics, Rutgers-New Jersey Medical School, Rutgers, The State University of New Jersey, Newark, NJ, USA.
Vivian
V
Bellofatto
Department of Medicine, Rutgers-New Jersey Medial School, OIT-High Performance and Research Computing, Rutgers, The State University of New Jersey, Newark, NJ, USA; Department of Invertebrate Zoology, American Museum of Natural History, New York, NY, USA.
Jeffrey
J
Rosenfeld
Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, UK.
Mark
M
Carrington
CITIC-UGR, Centro de Investigación en Tecnologías de la Información y de las Comunicaciones de la Universidad de Granada, Granada, Spain.
Rocío
R
Romero-Zaliz
CITIC-UGR, Centro de Investigación en Tecnologías de la Información y de las Comunicaciones de la Universidad de Granada, Granada, Spain.
Coral
C
del Val
Instituto de Parasitología y Biomedicina "López-Neyra", IPBLN-CSIC. Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento, s/n. 18016, Armilla, Granada, Spain. Electronic address: aestevez@ipb.csic.es.
Antonio M
AM
Estévez
D059014:High-Throughput Nucleotide Sequencing; D047468:Immunoprecipitation; D011485:Protein Binding; D015800:Protozoan Proteins / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D014346:Trypanosoma brucei brucei / Q000235:genetics*
D015800:Protozoan Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins
RNA-binding proteins; Trypanosoma brucei; mRNA translation