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Infection and Immunity, May 2003, p. 2555-2562, Vol. 71, No. 5
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.5.2555-2562.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Temporal Expression of Type III Secretion Genes of Chlamydia pneumoniae

Anatoly Slepenkin,1 Vladimir Motin,2 Luis M. de la Maza,1 and Ellena M. Peterson1*

Department of Pathology, University of California, Irvine, Irvine,1 Lawrence Livermore National Laboratory, Livermore, California2

Received 19 November 2002/ Returned for modification 14 January 2003/ Accepted 24 February 2003

Chlamydia pneumoniae has been shown to possess at least 13 genes that are homologous with other known type III secretion (TTS) systems. Upon infection of HEp-2 cells with C. pneumoniae, the expression of these genes was followed by reverse transcriptase PCR throughout the developmental cycle of this obligate intracellular pathogen. In addition, expression was analyzed when C. pneumoniae was grown in the presence of human gamma interferon (IFN-{gamma}). The groEL-1, ompA, and omcB genes were used as markers for the early, middle, and late stages of the developmental cycle, respectively, and the inhibition of expression of the fstK gene was used as a marker for the effect of IFN-{gamma} on the maturation of C. pneumoniae. In the absence of IFN-{gamma}, the TTS genes were expressed as follows: early stage (1.5 to 8 h), yscC, yscS, yscL, yscJ and lcrH-2; middle stage (by 12 to 18 h), lcrD, yscN, and yscR; and late stage (by 24 h), lcrE, sycE, lcrH-1, and yscT. Of the genes expressed early, the lcrH-2 gene was detected the earliest, at 1.5 h. Expression of the yscU gene was not detected at any of the time points examined. Under the influence of IFN-{gamma}, the cluster of TTS genes that were normally not expressed until the middle to late stages of the developmental cycle, namely, lcrD, lcrE, and sycE, as well as lcrH-1, were down-regulated, and expression could not be detected up to 48 h. In contrast, the expression of the other TTS genes appeared to be unchanged in the presence of IFN-{gamma}. The lcrH-1 and lcrH-2 genes differed from one another in both their temporal expression and response to IFN-{gamma}. In other TTS systems, these genes code for proteins that function in regulation of effector protein synthesis as well as serve as chaperones for proteins that provide for the translocation of the effector proteins into the host cell. In summary, the expression pattern of the TTS genes of C. pneumoniae examined suggests that they are temporally regulated throughout the developmental cycle. Furthermore, paralleling the inhibition of the maturation of the reticulate body to the elementary body, TTS genes expressed in the later stages of the cycle appear to be down-regulated when the organism is grown in the presence of IFN-{gamma}.

* Corresponding author. Mailing address: Department of Pathology, Medical Science Building, Room D-440, University of California Irvine, Irvine, CA 92697-4800. Phone: (949) 824-4169. Fax: (949) 824-2160. E-mail: epeterso{at}uci.edu.

Editor: D. L. Burns

Infection and Immunity, May 2003, p. 2555-2562, Vol. 71, No. 5
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.5.2555-2562.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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