Intracellular steady-state concentration of integron recombination products varies with integrase level and growth phase.
IntI1 mediates the recombination of antibiotic-resistant gene cassettes between different integrons in the same cell, facilitating the persistence and dissemination of these genes. Historically, integrase activity has been measured by conjugating recombinant products from donor cells overexpressing integrase and quantifying them in recipient cells. Here we report the first measurements of the steady-state intracellular abundance of integrase-mediated recombination products in strains expressing natural or high IntI1 levels. Recombination products in both high and natural integrase strains increased markedly through late log phase and continued to rise in stationary phase in the high integrase strain, but declined in the natural expression strain. Simple acquisition of gene cassettes was seen only in strains expressing high integrase; in strains with natural integrase levels, only cointegrates between the two integron-bearing plasmids were detectable at all growth stages. Unexpectedly, more attIxattI than attCxattI recombination products were seen in log phase for both strains; however, in stationary phase, the high integrase strain increased attC recombination, consistent with earlier observations of integrase crossover site preferences. Thus, direct quantification of the steady-state concentration of recombination products reveals that the integrase's intracellular concentration affects the amount and type of recombination events in a growth-phase-dependent manner.
Shearer
Julia E S
JE
Department of Microbiology, The University of Georgia, Athens, 30602-2605, USA.
Summers
Anne O
AO
D018407:Bacterial Physiological Phenomena; D002453:Cell Cycle; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D019426:Integrases / Q000378:metabolism*; D032023:Integrons; D011995:Recombination, Genetic
D004269:DNA, Bacterial; D019426:Integrases