Ribonucleoside-diphosphate reductase (RNR) is the sole source of de novo deoxyribonucleotide biosynthesis in almost all living organisms. Class I RNRs rely on metallocofactors for radical initiation and can be further divided into 3 subgroups (Ia, Ib and Ic) depending on the specific metal species and coordinating amino acid pattern. We recently identified a potentially novel class Ie subgroup via bioinformatics. This subgroup lacks 3 of the 6 conserved metal coordinating residues compared to canonical subgroups. In order to facilitate the investigation of its chemical mechanism, we introduced single mutations at selected sites around the proposed active center. Variant fitness was evaluated by applying selective pressure in an auxotrophic strain and deep-sequencing the variant populations before and after selection.
Submitter: Kai Hu
Submission Date: Oct. 1, 2018, midnight
|Number of data points||1340|
|Proteins||Ribonucleoside-diphosphate reductase, beta subunit|
|Assays/Quantities/Protocols||Experimental Assay: Sequencing read counts for unselected AuNrdF* variant library ; Experimental Assay: Sequencing read counts for selected AuNrdF* variant library ; Derived Quantity: Statistical analysis of read counts using Enrich2|
|Libraries||Variant fitness landscape|