Format

Send to

Choose Destination
Nucleic Acids Res. 2016 Apr 7;44(6):2962-73. doi: 10.1093/nar/gkw062. Epub 2016 Feb 2.

The oligomeric architecture of the archaeal exosome is important for processive and efficient RNA degradation.

Author information

1
Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 Tübingen, Germany.
2
Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 Tübingen, Germany remco.sprangers@tuebingen.mpg.de.

Abstract

The exosome plays an important role in RNA degradation and processing. In archaea, three Rrp41:Rrp42 heterodimers assemble into a barrel like structure that contains a narrow RNA entrance pore and a lumen that contains three active sites. Here, we demonstrate that this quaternary structure of the exosome is important for efficient RNA degradation. We find that the entrance pore of the barrel is required for nM substrate affinity. This strong interaction is crucial for processive substrate degradation and prevents premature release of the RNA from the enzyme. Using methyl TROSY NMR techniques, we establish that the 3' end of the substrate remains highly flexible inside the lumen. As a result, the RNA jumps between the three active sites that all equally participate in substrate degradation. The RNA jumping rate is, however, much faster than the cleavage rate, indicating that not all active site:substrate encounters result in catalysis. Enzymatic turnover therefore benefits from the confinement of the active sites and substrate in the lumen, which ensures that the RNA is at all times bound to one of the active sites. The evolution of the exosome into a hexameric complex and the optimization of its catalytic efficiency were thus likely co-occurring events.

PMID:
26837575
PMCID:
PMC4824110
DOI:
10.1093/nar/gkw062
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Silverchair Information Systems Icon for PubMed Central
Loading ...
Support Center