Willkomm, Sarah and Zander, Adrian and Grohmann, Dina and Restle, Tobias (2016) Mechanistic Insights into Archaeal and Human Argonaute Substrate Binding and Cleavage Properties. PLOS ONE, 11 (10): e0164695. ISSN 1932-6203,
Full text not available from this repository. (Request a copy)Abstract
Argonaute (Ago) proteins from all three domains of life are key players in processes that specifically regulate cellular nucleic acid levels. Some of these Ago proteins, among them human Argonaute2 (hAgo2) and Ago from the archaeal organism Methanocaldococcus jannaschii (MjAgo), are able to cleave nucleic acid target strands that are recognised via an Ago-associated complementary guide strand. Here we present an in-depth kinetic side-by-side analysis of hAgo2 and MjAgo guide and target substrate binding as well as target strand cleavage, which enabled us to disclose similarities and differences in the mechanistic pathways as a function of the chemical nature of the substrate. Testing all possible guide-target combinations (i.e. RNA/RNA, RNA/DNA, DNA/RNA and DNA/DNA) with both Ago variants we demonstrate that the molecular mechanism of substrate association is highly conserved among archaeal-eukaryotic Argonautes. Furthermore, we show that hAgo2 binds RNA and DNA guide strands in the same fashion. On the other hand, despite striking homology between the two Ago variants, MjAgo cannot orientate guide RNA substrates in a way that allows interaction with the target DNA in a cleavage-compatible orientation.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | RISC-LOADING COMPLEX; RNAI ENZYME COMPLEX; DOUBLE-STRANDED-RNA; CRYSTAL-STRUCTURE; GUIDE-STRAND; SILENCING COMPLEX; STRUCTURAL BASIS; ARCHAEOGLOBUS-FULGIDUS; AEOLICUS ARGONAUTE; PASSENGER-STRAND; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie > Prof. Dr. Dina Grohmann |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 24 Apr 2019 10:38 |
| Last Modified: | 24 Apr 2019 10:38 |
| URI: | https://pred.uni-regensburg.de/id/eprint/4099 |
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