Phenotypic Adaption of Pseudomonas aeruginosa by Hacking Siderophores Produced by Other Microorganisms

Abstract

Bacteria secrete siderophores to access iron, a key nutrient poorly bioavailable and the source of strong competition between microorganisms in most biotopes. Many bacteria also use siderophores produced by other microorganisms (exosiderophores) in a piracy strategy. Pseudomonas aeruginosa, an opportunistic pathogen, produces two siderophores, pyoverdine and pyochelin, and is also able to use a panel of exosiderophores. We first investigated expression of the various iron-uptake pathways of P. aeruginosa in three different growth media using proteomic and RT-qPCR approaches and observed three different phenotypic patterns, indicating complex phenotypic plasticity in the expression of the various iron-uptake pathways. We then investigated the phenotypic plasticity of iron-uptake pathway expression in the presence of various exosiderophores (present individually or as a mixture) under planktonic growth conditions, as well as in an epithelial cell infection assay. In all growth conditions tested, catechol-type exosiderophores were clearly more efficient in inducing the expression of their corresponding transporters than the others, showing that bacteria opt for the use of catechol siderophores to access in the In expression of the of the pathway under most conditions tested, as well as that of of the pyoverdine to a the expression of the and pathways. P. aeruginosa the expression of various iron-uptake pathways plasticity and to of and Bacteria secrete siderophores to access iron, a key nutrient poorly bioavailable and the source of strong competition between microorganisms in most biotopes. Many bacteria also use siderophores produced by other microorganisms (exosiderophores) in a piracy strategy. Pseudomonas aeruginosa, an opportunistic pathogen, produces two siderophores, pyoverdine and pyochelin, and is also able to use a panel of exosiderophores. We first investigated expression of the various iron-uptake pathways of P. aeruginosa in three different growth media using proteomic and RT-qPCR approaches and observed three different phenotypic patterns, indicating complex phenotypic plasticity in the expression of the various iron-uptake pathways. We then investigated the phenotypic plasticity of iron-uptake pathway expression in the presence of various exosiderophores (present individually or as a mixture) under planktonic growth conditions, as well as in an epithelial cell infection assay. In all growth conditions tested, catechol-type exosiderophores were clearly more efficient in inducing the expression of their corresponding transporters than the others, showing that bacteria opt for the use of catechol siderophores to access in the In expression of the of the pathway under most conditions tested, as well as that of of the pyoverdine to a the expression of the and pathways. P. aeruginosa the expression of various iron-uptake pathways plasticity and to of and all in competition for and is a key nutrient that is for and growth is in of and is the source of competition between in the and is a is poorly in under conditions and and competition for also in between and In competition for as well between and nutrient to by and of in the and of In a of competition for iron, bacteria to access the most the and of siderophores their and of a to and produced by bacteria under the by transporters of of and in the by bacteria to access to of different observed in the or the or siderophores and a corresponding for the of the and and of P. Pseudomonas aeruginosa in of the of In most bacteria able to also use exosiderophores produced by other able to the corresponding the and of the and and of P. Pseudomonas aeruginosa in of the of by and is a strong between the of various siderophores and exosiderophores that by a and the of in and and of transporters of Pseudomonas aeruginosa pyoverdine transporters of Pseudomonas aeruginosa pyoverdine competition for two use of and their in of as in and and competition in for and and a of Pseudomonas aeruginosa in the Pseudomonas the growth of in is a cell and cell between and in the Pseudomonas In bacteria siderophores and to siderophores Pseudomonas aeruginosa bacteria and the of their for growth under competition and in Pseudomonas and competition for in and Pseudomonas in of as a and of competition in the Pseudomonas aeruginosa Pseudomonas aeruginosa and to competition for In the the of the various siderophores produced the the bacteria the the for the to and access P. aeruginosa, an bacteria of different iron-uptake iron-uptake three iron-uptake pathways by the two siderophores, pyoverdine and produced by the pathogen, and different to using exosiderophores P. Pseudomonas aeruginosa in of the of a piracy is of the of P. aeruginosa to various pathways P. of in and the expression of the most efficient for the of Pseudomonas aeruginosa P. and cell and P. in and P. aeruginosa the presence of exosiderophores in using and two and a of the of Pseudomonas aeruginosa P. and cell and P. in and of the of Pseudomonas of a a of expression in Pseudomonas aeruginosa, as an and as a by the of Pseudomonas aeruginosa the as an in the presence of the of the as well as the to of the in the bacteria P. in and by the of Pseudomonas aeruginosa the as an siderophores the pathway and the pathway in Pseudomonas an for key for the in the in Pseudomonas in in Pseudomonas is piracy and competition in and the phenotypic bacteria and the expression of their various iron-uptake pathways in to is the expression of various pathways is that the bacteria to to and the various iron-uptake pathways in and their of expression to the of the phenotypic the expression of iron-uptake pathways growth conditions and investigated phenotypic plasticity in P. aeruginosa using proteomic and RT-qPCR We the of expression of various iron-uptake pathways in to different growth conditions growth in different media and an epithelial infection in the presence and of different exosiderophores a catechol produced by an for catechol produced by in a produced by of the and of in and produced by and in siderophores were than to the of the exosiderophores other the plasticity of the phenotypic In the of competition other siderophores, exosiderophores were all able to the expression of their corresponding pathways in P. aeruginosa and that the to P. aeruginosa In the presence of a of siderophores in the the of the various siderophores for a key and catechol siderophores strong the the in competition by other catechol siderophores the most efficient in inducing the expression of their corresponding in the presence of other siderophores under growth conditions, as well as in more complex as an epithelial cell infection assay. in of the of the pathway in conditions, also that of the of the the of expression of and pathways. and as Pseudomonas produces the of the Pseudomonas aeruginosa of pyochelin, a of Pseudomonas as P. and of siderophores of Pseudomonas aeruginosa and were and and P. aeruginosa in in in P. aeruginosa were first in and were then and in or were as the for the P. aeruginosa for were the of in of the and a the of the by using and a using to in the of P. aeruginosa were as by using P. aeruginosa and for by and key for the in the in Pseudomonas epithelial were in and to for the infection were and were between to cell using the cell were a of cell well in cell and to P. aeruginosa to an of in and for of growth in well of the and were of of siderophores bacteria of infection of In the a were also to by a the to and in a of a of were of a of for and and for of Bacteria were in in the presence of of the different exosiderophores in to the corresponding pathways. bacteria were then to the siderophores to transporters and to an of Bacteria were in the presence of bacteria were by and the in the were the the cell of the in Pseudomonas aeruginosa is to the to the of or of to the cell the in the presence of the in the of the to two siderophores were in competition in the siderophores were for and and in the of in the presence of then as using or siderophores were in in and or and in in of in the presence of of siderophores and for corresponding to for in a the of in of in the presence of siderophores and in of in a the using the to for the and of of were in of of of a of were in of and the then to by of of and of the to a of the of the in the presence of of siderophores and for for of in a expression by as siderophores the pathway and the pathway in Pseudomonas an for of by Pseudomonas of P. aeruginosa in were and in to an of were then in the presence or of or for of to two of Bacteria a and an We then of a in the of were in a and the the as an for a in were to for and as a two of the to the were a of cell in infection P. aeruginosa P. aeruginosa in under to an of and for the of the to indicating that bacteria in of were a of P. aeruginosa to of infection of were to the the as P. aeruginosa infection to for the were and the and bacteria to the of the a cell cell and bacteria were then by and the in of and two of Bacteria and the of the using were using the and then and a then as for the of in bacteria in the the of as a in to for of the of cell in in to an of were then or or for of were for proteomic in for cell were in of and and to efficient cell of of by of were by using as were in and the were in and under were in of and to and were of proteomic were by the of of a to an as P. P. is in were proteomic were the Pseudomonas aeruginosa a Pseudomonas aeruginosa and were were as two were and of were as and of as for and for were and for the and of to and were to and and of to three by all of all the and and the by different and a is also for in the and a of the the were to a using an the in to by an by as well as a were as for the RT-qPCR of and bacteria were in by for in of in were using the and of were a as a and for the were and were a for were using and for the were in of in and to the between were a to a as a and for an to in then to in and a of were were an and Pseudomonas aeruginosa and were to the and the of all as as and of as for and for were and for the and of to and were to and and the were using as for the for the of were using as and as were and were for the of were the to and the of a by P. aeruginosa We observed in the Pseudomonas between three or three for the in the or the presence of or using the to in in were using for all the for observed and for in to between two the using the were as corresponding to a between were for for for and for in growth media of Bacteria were in the conditions as for the RT-qPCR and proteomic growth media by were and in a in media using a We first investigated the of the various exosiderophores in to P. aeruginosa in the of competition other siderophores or using a and P. aeruginosa under conditions of Pseudomonas a for in the presence of of the exosiderophores to the expression of the of the corresponding iron-uptake of Pseudomonas aeruginosa Bacteria were then the and the bacteria of and were of and for two of siderophores of two siderophores a the of for for and for the other siderophores, We observed for that P. aeruginosa access the conditions a of the of the various transporters and their expression in P. aeruginosa in or were P. aeruginosa in or the to the in the P. aeruginosa in the media by proteomic in and in the of for in P. aeruginosa in in a by in the presence of various exosiderophores. were in to the expression of corresponding transporters of Pseudomonas aeruginosa Bacteria were then and by the corresponding to a of as in and were and the in the were also for in the presence of the to the of of to the cell or to were in the of to the of as of by or in in the presence of as a for were is to were by and or and were for in the presence of the various Bacteria were then by and the in panel in the presence of to the of that of of the to the cell and were in the of by or in in the presence of as a for were the is to were by and or and were as in or conditions in siderophores for the in as more the of for and to other We investigated by or two siderophores in the presence of or of and for and or or as a for and were to or under to or and and also to and in Pseudomonas pathway in Pseudomonas aeruginosa an of the and a of the and as for and were to the by or In the presence of the of by more than by We in the presence of as for by and by presence of and more the of to than the of two siderophores for and for of of a of Pseudomonas and a of Pseudomonas of the and that P. aeruginosa is able to access using various siderophores, the of of their to various siderophores in all the and different and is a strong of a of of a of Pseudomonas of the and siderophores in the in the presence of other siderophores, for the of various siderophores to by their to and the produced by P. aeruginosa is in of and and to a strong an for of of of a of Pseudomonas between and in Pseudomonas aeruginosa to different a of of for in the presence of of siderophores in of by the catechol-type siderophores and of We of by and the to to the of were also by of in the presence of siderophores were the siderophores and the for the of siderophores to siderophores clearly more for than the other siderophores or siderophores as and the of for We also a competition using In by and the complex a as an for the and of to a strong the the siderophores were able to and for were a for than for In in the presence of siderophores that catechol more efficient than the in and in competition for as the presence of catechol in the of P. aeruginosa the of to the bacteria and the of the different pathways poorly and siderophores produced of Pseudomonas a for of P. aeruginosa in corresponding to in indicating that is by the bacteria in that of the other were also in of Pseudomonas a for corresponding to and corresponding to the siderophores and and in in catechol-type siderophores, two of and and as in of the in expression between P. aeruginosa in in expression of the of the and iron-uptake pathways and that of the and in in the epithelial cell infection all the in bacteria in to as well In also and in by catechol-type in and of that transporters also under growth of in expression between P. aeruginosa in in the expression of of and and of in expression between P. aeruginosa in in in the expression of most of the to in in than in in the and and in by is under the of the and than that in In observed three different phenotypic for the expression of the in the of P. aeruginosa the were in three different and phenotypic is well that bacteria the presence of exosiderophores in their two or of Pseudomonas aeruginosa P. and cell and P. in and of the of Pseudomonas of a a of expression in Pseudomonas aeruginosa, as an and as a by the of Pseudomonas aeruginosa the as an P. aeruginosa in or of of the exosiderophores and a proteomic to the their expression or In a RT-qPCR to the proteomic the RT-qPCR an of the and expression of the is as well an of the and expression of the transporters or to the the in the pathway key for the in the in Pseudomonas or the in the pathway cell of the pathway in Pseudomonas proteomic that of in the growth of P. aeruginosa the expression of the of and the in the to siderophores the pathway and the pathway in Pseudomonas an for key for the in the in Pseudomonas by a in of presence of the other catechol in and expression of two and and a of of the than that observed for and for and to the and is the of and of and of siderophores produced by that a and or of Pseudomonas aeruginosa is for in Pseudomonas expression of by the presence of in and for the growth conditions in the presence of also an of the expression of a to P. aeruginosa and for an the and for of Pseudomonas in the Pseudomonas and of and to and an and to and for of Pseudomonas in the Pseudomonas and in the of the and in in P. the presence of the and expression of and to of the bacteria in the of pathway in Pseudomonas aeruginosa an of the and a of the that the in observed for in the presence of to that of in the presence of and is a catechol and able to P. aeruginosa also the of a in and that of other in and also in and of of the in of in the presence of different exosiderophores that catechol siderophores and more efficient than and in inducing the of their corresponding of expression by the presence of and that a for siderophores than proteomic and also that two catechol siderophores and or the expression of of the pathway others, the of the of the exosiderophores expression of the of the pathway under the growth conditions that is produced in to for of produced in the presence of the presence of and able to the of their corresponding In observed the of of the and pathways. expression of other and for their expression in by the presence of the exosiderophores In all exosiderophores were able to the and expression of and in the catechol siderophores the and expression of as well as all of the the expression of the of the the and or for other P. aeruginosa were in the presence of a of all exosiderophores two different and of in a of siderophores of and and the of the for in by RT-qPCR also of two different of for the proteomic in and We observed strong of the of the three in catechol and of for and and of for and of the the of of showing that siderophores, as in the presence of catechol-type siderophores and the expression of their corresponding were in the of three between and of the of more by of siderophores, in the presence of of than by the siderophores of and in the of the than is in the the that by to a in of and in the of a other than a observed of of indicating that is for to and the catechol of of of than as and in between and of In P. aeruginosa the expression of their iron-uptake pathways in the presence of of siderophores, two catechol siderophores, by the and expression of the catechol the expression of the pathway the and inducing the of in growth in We also investigated the to the presence of exosiderophores also in the epithelial cell infection assay. were P. aeruginosa in cell in to of the infection Bacteria the epithelial were of of the epithelial to that and the proteomic in infection were different for bacteria under planktonic growth conditions and to P. aeruginosa in the presence of a of the epithelial of the proteomic were to a to the expression of between bacteria in and the in of in P. aeruginosa for in in the or presence of epithelial by RT-qPCR in the P. aeruginosa were of presence of epithelial of and of and as well as of and of and two for that bacteria the presence of epithelial and the expression of their In also RT-qPCR to the of in in bacteria in and then for in bacteria in the conditions in in and in of all under the infection conditions of and for and the planktonic growth conditions in iron-uptake pathways of bacteria were of in epithelial to bacteria in were in use in the epithelial cell infection were as in as bacteria in in and for in RT-qPCR infection then or or siderophores were to the infection the as the bacteria in the bacteria were in the exosiderophores the of of and were and expression by proteomic and the of a investigated by RT-qPCR and in the in the of and by RT-qPCR of in the of in pathways in P. aeruginosa epithelial cell in the and presence of exosiderophores. RT-qPCR P. aeruginosa of in epithelial and or or RT-qPCR P. aeruginosa of epithelial in or or were to the and of three in all the as the between the in the presence of siderophores in their RT-qPCR were P. aeruginosa of epithelial in or a of and of a of of of as an and a for panel the were to the and of three in as the between the in the presence of siderophores in their and the iron-uptake pathways of P. aeruginosa in the infection were a than in bacteria in the bacteria were all exosiderophores the and expression of their the infection presence of the and expression of and of under conditions, and showing that the of exosiderophores to the expression of their corresponding also in complex as epithelial cell infection assay. the of and in in the presence of clearly in the of two different that observed of the of and a to P. aeruginosa the were also different observed in strong of and expression of in the epithelial infection expression in and in the of of or other iron-uptake pathways in the presence of and of the observed of of the and iron-uptake pathways in the in the of between and for and between and and to more access to the of their corresponding in the presence of than in their the as an nutrient epithelial cell bacteria in a in the presence of of the and expression of iron-uptake pathways to exosiderophores siderophores in the growth In the of the of an for and in of the of pathways. We also investigated the of the of or of the siderophores, and the of P. aeruginosa iron-uptake pathways the bacteria of the exosiderophores the of and as in that of or that of or and also were presence of of exosiderophores poorly to and the presence of epithelial as an In phenotypic plasticity for the and expression of iron-uptake pathways. P. aeruginosa and the expression of their iron-uptake pathways as a of presence of exosiderophores the and expression of their corresponding in an infection in bacteria poorly a of a in P. aeruginosa growth and as and expression in bacteria in planktonic conditions, in and of and key the the and and all and in in as and or and for more than the In is the of strong competition and the to siderophores a in the of bacteria to access nutrient and in of to a indicating that bacteria in the presence of siderophores, produced by other microorganisms of that most bacteria siderophores and their that transporters in the of In the to siderophores produced by other bacteria is to P. aeruginosa is presence of pathways in the that all In to the of by the phenotypic plasticity to the expression of iron-uptake pathways. P. aeruginosa is able to iron-uptake three iron-uptake pathways by the two siderophores and produced by the pathogen, and different to using exosiderophores P. Pseudomonas aeruginosa in of the of presence of a panel of pathways in the a panel of various of expression of iron-uptake the of P. aeruginosa in three different media and clearly three different phenotypic for the expression of the various iron-uptake pathways in the of pathways were in in in the of the expression of the and as well as the pathways and two transporters of to the in and in in a more complex in of iron-uptake pathway expression than for bacteria in the iron-uptake pathways as in were the of the expression of three other in and also that bacteria pathway a of a the to the growth Bacteria first in and then for in in the presence of of all to bacteria in as P. aeruginosa of in the presence of epithelial more the of for indicating that the bacteria the presence of the epithelial also that the the more the and expression of the various iron-uptake pathways than the presence of epithelial presence of exosiderophores or the expression of of the growth conditions and as well as other in the siderophores the in the presence of In the bacteria the presence of the in their and their to access the the more iron-uptake pathways to the of of exosiderophores in the of a the the of of their of the bacteria to and in different siderophores P. aeruginosa in the presence of a of the siderophores, the of of expression of the corresponding by the of the siderophores for iron, in the catechol-type siderophores the to their corresponding pathways and and siderophores, as in the presence of siderophores and inducing the expression of their corresponding epithelial cell infection also that the presence of the of exosiderophores to the of their corresponding pathways key in competition for clearly in the of the various siderophores to other in the the of expression of the of the various iron-uptake pathways in the Bacteria that use catechol siderophores an other most of the conditions catechol siderophores the and expression of the of the pathway as well as that of the to a by the of than for and for of a of Pseudomonas and a of Pseudomonas of the and that is first produced and is by the of between iron-uptake bacteria in a a for is to the of that P. aeruginosa in produces of than of Pseudomonas a for is in and in the presence of produced and in in the presence of to and for and the expression of the of the that in the presence of catechol siderophores a strong for for of P. aeruginosa first more able to the and the of bacteria to of a to in the of in the of of Pseudomonas a for is also an for the expression of a We observed of the of the and pathways in the presence of exosiderophores under of the growth conditions also of and expression under growth conditions tested, indicating that than and that transporters a in under growth a as is to P. aeruginosa in the also clearly the expression of various iron-uptake pathways and phenotypic the and complex phenotypic plasticity for the expression of the iron-uptake pathways in the P. aeruginosa for the of the bacteria to different that the siderophores competition for iron, and pathway in Pseudomonas aeruginosa an of the and a of the and of in the of in of the transporters in Bacteria a of their than for of the of a is in of the in Pseudomonas aeruginosa is is as the between the of a in the and the the the the to the for of to of of the and Bacteria more than to a of the a to a the of the expression of a iron-uptake pathway of the corresponding complex by the as well as of a of to their corresponding the In the and bacteria the expression of the iron-uptake pathways in their to their growth the expression of iron-uptake pathways. phenotypic plasticity and and by the and the presence of exosiderophores. siderophores, siderophores clearly the most a strong to the expression of their corresponding iron-uptake pathways in in bacteria that and in the competition for in of the and expression of the pathways is observed in the presence of siderophores, indicating that the bacteria use the siderophores to access iron, than their phenotypic plasticity the various iron-uptake pathways in P. aeruginosa a of to a of biotopes. of that of by siderophores an in the of the to to the bacteria as for Pseudomonas phenotypic plasticity of P. aeruginosa and that of other bacteria to a for the of a strategy. the strong of siderophores to the of their corresponding in various growth and in the presence of other siderophores, is a that and in were to the the the of the and and and

Topics & Concepts