Expression and localization of type III secretion-related proteins of Chlamydia pneumoniae.
The entire developmental cycle of the obligate intracellular bacteria Chlamydia pneumoniae takes place within the inclusion body. As many gram negative bacteria, Chlamydia possess a type III-secretion system (TTSS), which allows them to target effector molecules into the host cell. The expression and localization of several proteins constituting the TTSS apparatus and of proteins supposed to be secreted by the TTSS have been investigated. For the TTSS-constituting proteins, we selected representatives such as YscN (ATPase), LcrE (putative "lid" of the TTSS) and LcrH1 (postulated to be a chaperone). Furthermore, we focused on the putative effector proteins IncA, IncB, IncC, Cpn0809 and Cpn1020. Expression of these proteins was detected by reverse transcriptase-PCR followed by immunoblot analysis using antisera that were generated against the corresponding recombinant proteins. Thereby, expression could be detected on the RNA and/or protein level. Intracellular localization of proteins under investigation was determined by immunofluorescence assays using the respective antisera. YscN was shown to be distributed equally throughout the inclusion body, whereas LcrE gave a more prominent staining of the inclusion membrane. IncA was detected mainly on the membrane of the inclusion body, whereas IncB and IncC were shown to be located within the inclusion. Immunofluorescence assays with antisera raised against Cpn0809 and Cpn1020 showed completely different labeling. Signals corresponding to Cpn0809 and Cpn1020 were distributed within the host cell rather than inside the inclusions. Taken together, the different localization patterns of the effector proteins indicate differences in function and interplay with the host cell.
Department of Bacteriology, University of Göttingen, Kreuzbergring 57, 37075 Göttingen, Germany.
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R
Lugert
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M
Kuhns
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T
Polch
U
U
Gross
D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001692:Biological Transport; D015153:Blotting, Western; D002478:Cells, Cultured; D016993:Chlamydophila pneumoniae / Q000737:chemistry / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D012329:RNA, Bacterial; D012333:RNA, Messenger; D020133:Reverse Transcriptase Polymerase Chain Reaction
D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger