Temporal Expression of Type III Secretion Genes ofChlamydia pneumoniae

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Abstract
Chlamydia pneumoniaehas 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 withC. 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 whenC. pneumoniaewas grown in the presence of human gamma interferon (IFN-γ). ThegroEL-1,ompA, andomcBgenes were used as markers for the early, middle, and late stages of the developmental cycle, respectively, and the inhibition of expression of thefstKgene was used as a marker for the effect of IFN-γ on the maturation ofC. pneumoniae. In the absence of IFN-γ, the TTS genes were expressed as follows: early stage (1.5 to 8 h),yscC,yscS,yscL,yscJandlcrH-2; middle stage (by 12 to 18 h),lcrD,yscN, andyscR; and late stage (by 24 h),lcrE,sycE,lcrH-1, andyscT. Of the genes expressed early, thelcrH-2gene was detected the earliest, at 1.5 h. Expression of theyscUgene was not detected at any of the time points examined. Under the influence of IFN-γ, the cluster of TTS genes that were normally not expressed until the middle to late stages of the developmental cycle, namely,lcrD,lcrE, andsycE, as well aslcrH-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-γ. ThelcrH-1andlcrH-2genes differed from one another in both their temporal expression and response to IFN-γ. 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 ofC. pneumoniaeexamined 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-γ.