Regulated synthesis of the Borrelia burgdorferi inner-membrane lipoprotein IpLA7 (P22, P22-A) during the Lyme disease spirochaete's mammal-tick infectious cycle.
Results of previous immunological studies suggested that Borrelia burgdorferi regulates synthesis of the IpLA7 lipoprotein during mammalian infection. Through combined use of quantitative reverse transcription PCR, immunofluorescence analyses, ELISA and immunoblotting, it is now demonstrated that IpLA7 is actually expressed throughout mammalian infection, as well as during transmission both from feeding ticks to naïve mice and from infected mice to naïve, feeding ticks. However, proportions of IpLA7-expressing B. burgdorferi within tick midguts declined significantly with time following completion of blood feeding. Cultured bacteria differentially expressed IpLA7 in response to changes in temperature, pH and concentration of 4,5-dihydroxy-2,3-pentanedione, the precursor of autoinducer 2, indicative of mechanisms governing IpLA7 expression. Previous studies also reported mixed results as to the cellular localization of IpLA7. It is now demonstrated that IpLA7 localizes primarily to the borrelial inner membrane and is not surface-exposed, consistent with the ability of these bacteria to produce IpLA7 throughout mammalian infection despite being the target of a robust immune response.
von Lackum
Kate
K
Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
Ollison
Kristina M
KM
Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
Bykowski
Tomasz
T
Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
Nowalk
Andrew J
AJ
Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.|Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Hughes
Jessica L
JL
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Carroll
James A
JA
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Zückert
Wolfram R
WR
Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
Stevenson
Brian
B
Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D015153:Blotting, Western; D025065:Borrelia burgdorferi / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000737:chemistry; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D041981:Gastrointestinal Tract / Q000382:microbiology; D015964:Gene Expression Regulation, Bacterial; D006863:Hydrogen-Ion Concentration; D008074:Lipoproteins / Q000096:biosynthesis* / Q000235:genetics; D008193:Lyme Disease / Q000382:microbiology; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D010420:Pentanes / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature; D013987:Ticks / Q000382:microbiology
C493138:4,5-dihydroxy-2,3-pentanedione; D001426:Bacterial Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D010420:Pentanes; D012333:RNA, Messenger