Analyzing the variant fitness landscape of a novel ribonucleoside-diphosphate reductase by saturation mutagenesis and deep sequencing


Abstract

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.

Submission Details

ID: JchqkPwm

Submitter: Kai Hu

Submission Date: Feb. 14, 2018, 12:19 p.m.

Version: 1

Publication Details
Kai Hu, Amelia J Kim, Benjamin D Allen,unpublished (2018) AuNrdF_valid_input
Additional Information

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)