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Atypical roles for campylobacter jejuni AA-ABC transporter components PAQP and PAQQ in bacterial stress tolerance and pathogen-host cell dynamicsLin, Ann En-Ju 11 1900 (has links)
Campylobacter jejuni is a human pathogen that causes severe diarrhea! disease. However,
our understanding of C. jejuni virulence mechanisms and survival during disease and
transmission remains limited. Amino acid ATP Binding Cassette (AA-ABC) transporters in C.
jejuni have been proposed as being important for bacterial physiology and pathogenesis. We
have investigated a novel AA-ABC transporter system, encoded by cj0467-9, by generating
targeted deletions of cj0467 (membrane transport component) and cj0469 (ATPase component)
in C. jejuni 81-176. Analyses described herein have led us to designate these genes paqP and
paqQ, respectively [pathogenesis-ssociated glutamine (q) ABC transporter permease () and
ATPase (Q)]. We found that loss of either component resulted in amino acid uptake defects,
most notably diminished glutamine uptake. Both ΔpaqP and ΔpaqQ mutants also exhibited a
surprising but significant increase in short-term intracellular survival in macrophages and
epithelial cells. Levels of resistance to a series of environmental and in vivo stresses were
examined. Both mutants were hyper-resistant to aerobic and oxidative stress, and while ΔpaqP
was also hyper-resistant to heat and osmotic shock, ΔpaqQ was more susceptible than wild-type
to the latter two stresses. Annexin-V staining coupled with fluorescence microscopy revealed
that macrophages infected with the ΔpaqP and ΔpaqQ mutants underwent a lower level of
apoptosis than cells infected with wild-type bacteria. Macrophages infected with the mutant
strains exhibited a transient decrease in ERK activation compared to wild type-infected
macrophages, potentially explaining the reduced apoptosis phenotype. The ΔpaqP mutant did not
exhibit a defect for short or longer term mouse colonization, consistent with its increased stress
survival and diminished host cell damage phenotypes. Collectively, these results demonstrate a
unique correlation between an AA-ABC transporter with bacterial stress tolerance, intracellular
survival, host cell damage, and host signal transduction in response to pathogen infection.
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Atypical roles for campylobacter jejuni AA-ABC transporter components PAQP and PAQQ in bacterial stress tolerance and pathogen-host cell dynamicsLin, Ann En-Ju 11 1900 (has links)
Campylobacter jejuni is a human pathogen that causes severe diarrhea! disease. However,
our understanding of C. jejuni virulence mechanisms and survival during disease and
transmission remains limited. Amino acid ATP Binding Cassette (AA-ABC) transporters in C.
jejuni have been proposed as being important for bacterial physiology and pathogenesis. We
have investigated a novel AA-ABC transporter system, encoded by cj0467-9, by generating
targeted deletions of cj0467 (membrane transport component) and cj0469 (ATPase component)
in C. jejuni 81-176. Analyses described herein have led us to designate these genes paqP and
paqQ, respectively [pathogenesis-ssociated glutamine (q) ABC transporter permease () and
ATPase (Q)]. We found that loss of either component resulted in amino acid uptake defects,
most notably diminished glutamine uptake. Both ΔpaqP and ΔpaqQ mutants also exhibited a
surprising but significant increase in short-term intracellular survival in macrophages and
epithelial cells. Levels of resistance to a series of environmental and in vivo stresses were
examined. Both mutants were hyper-resistant to aerobic and oxidative stress, and while ΔpaqP
was also hyper-resistant to heat and osmotic shock, ΔpaqQ was more susceptible than wild-type
to the latter two stresses. Annexin-V staining coupled with fluorescence microscopy revealed
that macrophages infected with the ΔpaqP and ΔpaqQ mutants underwent a lower level of
apoptosis than cells infected with wild-type bacteria. Macrophages infected with the mutant
strains exhibited a transient decrease in ERK activation compared to wild type-infected
macrophages, potentially explaining the reduced apoptosis phenotype. The ΔpaqP mutant did not
exhibit a defect for short or longer term mouse colonization, consistent with its increased stress
survival and diminished host cell damage phenotypes. Collectively, these results demonstrate a
unique correlation between an AA-ABC transporter with bacterial stress tolerance, intracellular
survival, host cell damage, and host signal transduction in response to pathogen infection.
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Atypical roles for campylobacter jejuni AA-ABC transporter components PAQP and PAQQ in bacterial stress tolerance and pathogen-host cell dynamicsLin, Ann En-Ju 11 1900 (has links)
Campylobacter jejuni is a human pathogen that causes severe diarrhea! disease. However,
our understanding of C. jejuni virulence mechanisms and survival during disease and
transmission remains limited. Amino acid ATP Binding Cassette (AA-ABC) transporters in C.
jejuni have been proposed as being important for bacterial physiology and pathogenesis. We
have investigated a novel AA-ABC transporter system, encoded by cj0467-9, by generating
targeted deletions of cj0467 (membrane transport component) and cj0469 (ATPase component)
in C. jejuni 81-176. Analyses described herein have led us to designate these genes paqP and
paqQ, respectively [pathogenesis-ssociated glutamine (q) ABC transporter permease () and
ATPase (Q)]. We found that loss of either component resulted in amino acid uptake defects,
most notably diminished glutamine uptake. Both ΔpaqP and ΔpaqQ mutants also exhibited a
surprising but significant increase in short-term intracellular survival in macrophages and
epithelial cells. Levels of resistance to a series of environmental and in vivo stresses were
examined. Both mutants were hyper-resistant to aerobic and oxidative stress, and while ΔpaqP
was also hyper-resistant to heat and osmotic shock, ΔpaqQ was more susceptible than wild-type
to the latter two stresses. Annexin-V staining coupled with fluorescence microscopy revealed
that macrophages infected with the ΔpaqP and ΔpaqQ mutants underwent a lower level of
apoptosis than cells infected with wild-type bacteria. Macrophages infected with the mutant
strains exhibited a transient decrease in ERK activation compared to wild type-infected
macrophages, potentially explaining the reduced apoptosis phenotype. The ΔpaqP mutant did not
exhibit a defect for short or longer term mouse colonization, consistent with its increased stress
survival and diminished host cell damage phenotypes. Collectively, these results demonstrate a
unique correlation between an AA-ABC transporter with bacterial stress tolerance, intracellular
survival, host cell damage, and host signal transduction in response to pathogen infection. / Science, Faculty of / Microbiology and Immunology, Department of / Graduate
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