Spliceosomal Prp8 intein at the crossroads of protein and RNA splicing.

Published online

Journal Article

The spliceosome is a large ribonucleoprotein complex that removes introns from pre-mRNAs. At its functional core lies the essential pre-mRNA processing factor 8 (Prp8) protein. Across diverse eukaryotes, this protein cofactor of RNA catalysis harbors a self-splicing element called an intein. Inteins in Prp8 are extremely pervasive and are found at 7 different sites in various species. Here, we focus on the Prp8 intein from Cryptococcus neoformans (Cne), a human fungal pathogen. We solved the crystal structure of this intein, revealing structural homology among protein splicing sequences in eukaryotes, including the Hedgehog C terminus. Working with the Cne Prp8 intein in a reporter assay, we find that the biologically relevant divalent metals copper and zinc inhibit intein splicing, albeit by 2 different mechanisms. Copper likely stimulates reversible modifications on a catalytically important cysteine, whereas zinc binds at the terminal asparagine and the same critical cysteine. Importantly, we also show that copper treatment inhibits Prp8 protein splicing in Cne. Lastly, an intein-containing Prp8 precursor model is presented, suggesting that metal-induced protein splicing inhibition would disturb function of both Prp8 and the spliceosome. These results indicate that Prp8 protein splicing can be modulated, with potential functional implications for the spliceosome.

Full Text

Duke Authors

Cited Authors

  • Green, CM; Li, Z; Smith, AD; Novikova, O; Bacot-Davis, VR; Gao, F; Hu, S; Banavali, NK; Thiele, DJ; Li, H; Belfort, M

Published Date

  • October 2019

Published In

Volume / Issue

  • 17 / 10

Start / End Page

  • e3000104 -

PubMed ID

  • 31600193

Pubmed Central ID

  • 31600193

Electronic International Standard Serial Number (EISSN)

  • 1545-7885

Digital Object Identifier (DOI)

  • 10.1371/journal.pbio.3000104

Language

  • eng

Conference Location

  • United States