In order to predict whether organisms will evolve resistance to new antibiotics or whether organisms in live vaccines will re-evolve virulence we need to predict evolutionary changes that are the consequence of defined selections for new or improved biological functions. The proposed project seeks to answer the questions """"""""What information about a microorganism do we require in order to accurately predict which genes will mutate, and what mutations will occur, in order for that organism to evolve a new function in response to a specific selective pressure?"""""""" The model system will be the evolution of a Lac PTS/phospho-beta- galactosidase system for the catabolism of lactose in Escherichia coli. Two distinct functions are required: (1) an EIIlac for transport and phosphorylation of lactose and (2) a phospho-beta-galactosidase enzyme to cleave the phosphorylated lactose. There are five distinct gene systems, each of them for the catabolism of beta-glucoside sugars, that are good candidates to evolve the new lactose-specific functions. There are four different kinds of information that can be used to predict which genes, and which sites within those genes, will evolve: (1) functional comparison approach, (2) the sequence/phylogeny approach, (3) the biochemical approach. (4) the fitness measurement approach, (5) the in vitro sexual-PCR approach. First the data required to make predictions according to each of those approaches will be obtained for each of the four gene systems. It is likely that the EIIlac function and the phospho beta-galactosidase functions will evolve from different genes systems. Therefore strains that express each of the possible pairs of gene systems will be used to select spontaneous mutants that express the evolved Lac PTS/phospho-beta-galactosidase system. The resulting mutants will then be characterized to determine which kind of data most accurately predicted the eventual evolutionary outcome.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM060761-01
Application #
6053031
Study Section
Special Emphasis Panel (ZRG1-GEN (01))
Program Officer
Eckstrand, Irene A
Project Start
2000-01-01
Project End
2003-12-31
Budget Start
2000-01-01
Budget End
2000-12-31
Support Year
1
Fiscal Year
2000
Total Cost
$164,675
Indirect Cost
Name
University of Rochester
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
208469486
City
Rochester
State
NY
Country
United States
Zip Code
14627
Hall, Barry G (2004) In vitro evolution predicts that the IMP-1 metallo-beta-lactamase does not have the potential to evolve increased activity against imipenem. Antimicrob Agents Chemother 48:1032-3
Hall, Barry G; Salipante, Stephen J; Barlow, Miriam (2004) Independent origins of subgroup Bl + B2 and subgroup B3 metallo-beta-lactamases. J Mol Evol 59:133-41
Hall, Barry G; Salipante, Stephen J; Barlow, Miriam (2003) The metallo-beta-lactamases fall into two distinct phylogenetic groups. J Mol Evol 57:249-54
Salipante, Stephen J; Barlow, Miriam; Hall, Barry G (2003) GeneHunter, a transposon tool for identification and isolation of cryptic antibiotic resistance genes. Antimicrob Agents Chemother 47:3840-5
Hall, Barry G; Barlow, Miriam (2003) Structure-based phylogenies of the serine beta-lactamases. J Mol Evol 57:255-60
Salipante, Stephen J; Hall, Barry G (2003) Determining the limits of the evolutionary potential of an antibiotic resistance gene. Mol Biol Evol 20:653-9
Barlow, Miriam; Hall, Barry G (2003) Experimental prediction of the natural evolution of antibiotic resistance. Genetics 163:1237-41
Barlow, Miriam; Hall, Barry G (2003) Experimental prediction of the evolution of cefepime resistance from the CMY-2 AmpC beta-lactamase. Genetics 164:23-9
Barlow, Miriam; Hall, Barry G (2002) Predicting evolutionary potential: in vitro evolution accurately reproduces natural evolution of the tem beta-lactamase. Genetics 160:823-32
Hall, Barry G (2002) Predicting evolution by in vitro evolution requires determining evolutionary pathways. Antimicrob Agents Chemother 46:3035-8

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