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Verfasst von:Waithaka, Albina [VerfasserIn]   i
 Maiakovska, Olena [VerfasserIn]   i
 Grimm, Dirk [VerfasserIn]   i
 Melo do Nascimento, Larissa [VerfasserIn]   i
 Clayton, Christine [VerfasserIn]   i
Titel:Sequences and proteins that influence mRNA processing in Trypanosoma brucei
Titelzusatz:evolutionary conservation of SR-domain and PTB protein functions
Verf.angabe:Albina Waithaka, Olena Maiakovska, Dirk Grimm, Larissa Melo do Nascimento, Christine Clayton
Ausgabe:Version 2
E-Jahr:2022
Jahr:October 26, 2022
Umfang:31 S.
Fussnoten:Gesehen am 10.03.2023
Titel Quelle:Enthalten in: Public Library of SciencePLoS neglected tropical diseases
Ort Quelle:Lawrence, Kan. : PLoS, 2007
Jahr Quelle:2022
Band/Heft Quelle:16(2022), 10, Artikel-ID i0010876, Seite 1-31
ISSN Quelle:1935-2735
Abstract:Background Spliced leader trans splicing is the addition of a short, capped sequence to the 5’ end of mRNAs. It is widespread in eukaryotic evolution, but factors that influence trans splicing acceptor site choice have been little investigated. In Kinetoplastids, all protein-coding mRNAs are 5’ trans spliced. A polypyrimidine tract is usually found upstream of the AG splice acceptor, but there is no branch point consensus; moreover, splicing dictates polyadenylation of the preceding mRNA, which is a validated drug target. Methodology and principal findings We here describe a trans splicing reporter system that can be used for studies and screens concerning the roles of sequences and proteins in processing site choice and efficiency. Splicing was poor with poly(U) tracts less than 9 nt long, and was influenced by an intergenic region secondary structure. A screen for signals resulted in selection of sequences that were on average 45% U and 35% C. Tethering of either the splicing factor SF1, or the cleavage and polyadenylation factor CPSF3 within the intron stimulated processing in the correct positions, while tethering of two possible homologues of Opisthokont PTB inhibited processing. In contrast, tethering of SR-domain proteins RBSR1, RBSR2, or TSR1 or its interaction partner TSR1IP, promoted use of alternative signals upstream of the tethering sites. RBSR1 interacts predominantly with proteins implicated in splicing, whereas the interactome of RBSR2 is more diverse. Conclusions Our selectable constructs are suitable for screens of both sequences, and proteins that affect mRNA processing in T. brucei. Our results suggest that the functions of PTB and SR-domain proteins in splice site definition may already have been present in the last eukaryotic common ancestor.
DOI:doi:10.1371/journal.pntd.0010876
URL:Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.

kostenfrei: Volltext: https://doi.org/10.1371/journal.pntd.0010876
 kostenfrei: Volltext: https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0010876
 DOI: https://doi.org/10.1371/journal.pntd.0010876
Datenträger:Online-Ressource
Sprache:eng
Sach-SW:Cloning
 Messenger RNA
 Northern blot
 Polyadenylation
 RNA interference
 RNA splicing
 Transcriptome analysis
 Trypanosoma
K10plus-PPN:1838850295
Verknüpfungen:→ Zeitschrift

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