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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 107, No. 4, 2012, pp. 522-531
Bioline Code: oc12076
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 107, No. 4, 2012, pp. 522-531

 en Footprints of a trypanosomatid RNA world: pre-small subunit rRNA processing by spliced leader addition trans-splicing
Mayer, Mario Gustavo; dos Santos, Marcos Gonzaga; da Silva, Maria Fernanda Laranjeira & Floeter-Winter, Lucile Maria

Abstract

The addition of a capped mini-exon [spliced leader (SL)] through trans-splicing is essential for the maturation of RNA polymerase (pol) II-transcribed polycistronic pre-mRNAs in all members of the Trypanosomatidae family. This process is an inter-molecular splicing reaction that follows the same basic rules of cis-splicing reactions. In this study, we demonstrated that mini-exons were added to precursor ribosomal RNA (pre-rRNA) are transcribed by RNA pol I, including the 5’ external transcribed spacer (ETS) region. Additionally, we detected the SL-5’ETS molecule using three distinct methods and located the acceptor site between two known 5’ETS rRNA processing sites (A’ and A1) in four different trypanosomatids. Moreover, we detected a polyadenylated 5’ETS upstream of the trans-splicing acceptor site, which also occurs in pre-mRNA trans-splicing. After treatment with an indirect trans-splicing inhibitor (sinefungin), we observed SL-5’ETS decay. However, treatment with 5-fluorouracil (a precursor of RNA synthesis that inhibits the degradation of pre-rRNA) led to the accumulation of SL-5’ETS, suggesting that the molecule may play a role in rRNA degradation. The detection of trans-splicing in these molecules may indicate broad RNA-joining properties, regardless of the polymerase used for transcription.

Keywords
external transcribed spacer - spliced leader RNA - RNA polymerase I - spliceosome - RNA polymerase II carboxy-terminal domain

 
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