Capturing the mutational landscape of the beta-lactamase TEM-1


Abstract

Adaptation proceeds through the selection of mutations. The distribution of mutant fitness effect and the forces shaping this distribution are therefore keys to predict the evolutionary fate of organisms and their constituents such as enzymes. Here, by producing and sequencing a comprehensive collection of 10,000 mutants, we explore the mutational landscape of one enzyme involved in the spread of antibiotic resistance, the beta-lactamase TEM-1. We measured mutation impact on the enzyme activity through the estimation of amoxicillin minimum inhibitory concentration on a subset of 990 mutants carrying a unique missense mutation, representing 64% of possible amino acid changes in that protein reachable by point mutation. We established that mutation type, solvent accessibility of residues, and the predicted effect of mutations on protein stability primarily determined alone or in combination changes in minimum inhibitory concentration of mutants. Moreover, we were able to capture the drastic modification of the mutational landscape induced by a single stabilizing point mutation (M182T) by a simple model of protein stability. This work thereby provides an integrated framework to study mutation effects and a tool to understand/define better the epistatic interactions.

Submission Details

ID: LPbrW77i

Submitter: Connie Wang

Submission Date: Oct. 24, 2016, 4:40 p.m.

Version: 1

Publication Details
Jacquier H;Birgy A;Le Nagard H;Mechulam Y;Schmitt E;Glodt J;Bercot B;Petit E;Poulain J;Barnaud G;Gros PA;Tenaillon O,Proc Natl Acad Sci U S A (2013) Capturing the mutational landscape of the beta-lactamase TEM-1. PMID:23878237
Additional Information

Number of data points 990
Proteins beta-lactamase TEM-1
Unique complexes 990
Assays/Quantities/Protocols Experimental Assay: Survival in Amoxicillin
Libraries Mutational landscape of beta-lactamase
Sequence Assay Result Units