Capturing the Mutational Landscape of the Beta-lactamase TEM-1

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2013 Jul 24

PMID 23878237

Citations 128

Authors

Herve Jacquier

Andre Birgy

Herve Le Nagard

Yves Mechulam

Emmanuelle Schmitt

Jeremy Glodt

Beatrice Bercot

Emmanuelle Petit

Julie Poulain

Guilene Barnaud

Pierre-Alexis Gros

Olivier Tenaillon

Affiliations

Soon will be listed here.

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.

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