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Identification of iron-responsive proteins expressed by Chlamydia trachomatis reticulate bodies during intracellular growth

Brian D. Dill

Department of Microbiology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN 37614-1700, USA

Correspondence
Brian D. Dill
dillbd{at}ornl.gov

The obligate intracellular bacterium Chlamydia trachomatis serovar E is the most prevalent cause of bacterial sexually transmitted disease. With an established requirement for iron, the developmental cycle arrests at the intracellular reticulate body stage during iron restriction, resulting in a phenomenon termed persistence. Persistence has implications in natural infections for altered expression of virulence factors and antigens, in addition to a potential role in producing chronic infection. In this study, chlamydial proteins in iron-restricted, infected HEC-1B cells were radiolabelled during mid-developmental cycle growth, harvested, and separated using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Of 250 radiolabelled protein species visualized, densitometric analysis revealed 25 proteins that increased in expression under iron restriction compared to iron-sufficient control samples; ten protein species identified by mass spectrometry are involved in the oxidative damage response (alkyl hydroperoxide reductase, 6-phosphogluconolactonase and acyl carrier protein synthase), transcription (RNA polymerase subunit alpha and transcription anti-termination factors NusA and NusG), protein modification (peptide deformylase and trigger factor), and virulence (Chlamydia protein associating with death domains, CADD). Transcript-level expression patterns of ahpC, devB, cadd, fabF and ct538 were measured by quantitative RT-PCR throughout the developmental cycle, and each gene examined demonstrated a significant but small mid-cycle increase in transcript level in iron-restricted cultures compared to iron-replete controls. Taken together, these data suggest that the primary response of chlamydiae to reduced iron availability is to increase expression of proteins involved in protection against oxidative damage via iron-catalysed generation of reactive oxygen species and adaptation to stress by increasing expression of transcriptional machinery and other stress-responsive proteins.

Present Address: Organic and Biological Mass Spectrometry Group, Chemical Sciences Division, Oak Ridge National Laboratory, PO Box 2008, MS6131, Oak Ridge, TN 37831-6131, USA.

This article has been cited by other articles:

				J. D. Miller, M. S. Sal, M. Schell, J. D. Whittimore, and J. E. Raulston Chlamydia trachomatis YtgA is an iron-binding periplasmic protein induced by iron restriction Microbiology, September 1, 2009; 155(9): 2884 - 2894. [Abstract] [Full Text] [PDF]