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TitleStructural basis of Spliced Leader RNA recognition by the Trypanosoma brucei cap-binding complex.
Journal, issue, pagesNat Commun, Vol. 16, Issue 1, Page 685, Year 2025
Publish dateJan 15, 2025
AuthorsHarald Bernhard / Hana Petržílková / Barbora Popelářová / Kamil Ziemkiewicz / Karolina Bartosik / Marcin Warmiński / Laura Tengo / Henri Gröger / Luciano G Dolce / Cameron D Mackereth / Ronald Micura / Jacek Jemielity / Eva Kowalinski /
PubMed AbstractKinetoplastids are a clade of eukaryotic protozoans that include human parasitic pathogens like trypanosomes and Leishmania species. In these organisms, protein-coding genes are transcribed as ...Kinetoplastids are a clade of eukaryotic protozoans that include human parasitic pathogens like trypanosomes and Leishmania species. In these organisms, protein-coding genes are transcribed as polycistronic pre-mRNAs, which need to be processed by the coupled action of trans-splicing and polyadenylation to yield monogenic mature mRNAs. During trans-splicing, a universal RNA sequence, the spliced leader RNA (SL RNA) mini-exon, is added to the 5'-end of each mRNA. The 5'-end of this mini-exon carries a hypermethylated cap structure and is bound by a trypanosomatid-specific cap-binding complex (CBC). The function of three of the kinetoplastid CBC subunits is unknown, but an essential role in cap-binding and trans-splicing has been suggested. Here, we report cryo-EM structures that reveal the molecular architecture of the Trypanosoma brucei CBC (TbCBC) complex. We find that TbCBC interacts with two distinct features of the SL RNA. The TbCBP20 subunit interacts with the mG cap while TbCBP66 recognizes double-stranded portions of the SL RNA. Our findings pave the way for future research on mRNA maturation in kinetoplastids. Moreover, the observed structural similarities and differences between TbCBC and the mammalian cap-binding complex will be crucial for considering the potential of TbCBC as a target for anti-trypanosomatid drug development.
External linksNat Commun / PubMed:39814716 / PubMed Central
MethodsEM (single particle)
Resolution2.4 - 2.8 Å
Structure data

50173

9f3f

EMDB-50173, PDB-9f3f:
Trypanosoma brucei nuclear cap-binding complex (CBC) bound to cap0
Method: EM (single particle) / Resolution: 2.4 Å

50217

9f67

EMDB-50217, PDB-9f67:
Trypanosoma brucei nuclear cap-binding complex (CBC) bound to cap4
Method: EM (single particle) / Resolution: 2.8 Å

Chemicals

ChemComp-7MG:
7N-METHYL-8-HYDROGUANOSINE-5'-MONOPHOSPHATE

Source
  • trypanosoma brucei brucei (eukaryote)
  • synthetic construct (others)
KeywordsRNA BINDING PROTEIN / nuclear cap binding complex / CBC / cap0 / cap-4

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