Quantifying the Adaptive Potential of an Antibiotic Resistance Enzyme 英文参考文献.docVIP

QuantifyingtheAdaptivePotentialofanAntibiotic ResistanceEnzyme MartijnF.Schenk1,2,IvanG.Szendro3,JoachimKrug3,4,J.ArjanG.M.deVisser2* 1InstituteforGenetics,UniversityofCologne,Ko¨ln,Germany,2LaboratoryofGenetics,WageningenUniversity,Wageningen,TheNetherlands,3InstituteforTheoretical Physics,UniversityofCologne,Ko¨ln,Germany,4SystemsBiologyofAgeingCologne(Sybacol),UniversityofCologne,Ko¨ln,Germany Abstract For a quantitative understanding of the process of adaptation, we need to understand its ‘‘raw material,’’ that is, the frequency and fitness effects of beneficial mutations. At present, most empirical evidence suggests an exponential distribution of fitness effects of beneficial mutations, as predicted for Gumbel-domain distributions by extreme value theory.Here,westudythedistributionofmutationeffectsoncefotaxime(Ctx)resistanceandfitnessof48uniquebeneficial mutations in the bacterial enzyme TEM-1 b-lactamase, which were obtained by screening the products of random mutagenesisforincreasedCtxresistance.Ourcontributionsarethreefold.First,basedonthefrequencyofuniquemutations among more than 300 sequenced isolates and correcting for mutation bias, we conservatively estimate that the total numberoffirst-stepmutationsthatincreaseCtxresistanceinthisenzymeis87[95%CI75–189],or3.4%ofall2,583possible base-pairsubstitutions.Ofthe48mutations,10aresynonymousandthemajorityofthe38non-synonymousmutations occurinthepocketsurroundingthecatalyticsite.Second,weestimatetheeffectsofthemutationsonCtxresistanceby determining survival at various Ctx concentrations, and we derive their fitness effects by modeling reproduction and survivalasabranchingprocess.Third,wefindthatthedistributionofbothmeasuresfollowsaFre′chet-type distribution characterized by a broad tail of a few exceptionally fit mutants. Such distributions have fundamental evolutionary implications, including an increased predictability of evolution, and may provide a partial explanation for recent observationsofstrik