e Chlamydia trachomatis is the most prevalent cause of preventable blindness worldwide and a major reason for infectious infertility in females. Several bacterial factors have been implicated in the pathogenesis of C. trachomatis. Combining structural and mutational analysis, we have shown that the proteolytic function of CT441 depends on a conserved Ser/Lys/Gln catalytic triad and a functional substrate-binding site within a flexible PDZ (postsynaptic density of 95 kDa, discs large, and zonula occludens) domain. Previously, it has been suggested that CT441 is involved in modulating estrogen signaling responses of the host cell. Our results show that although in vitro CT441 exhibits proteolytic activity against SRAP1, a coactivator of estrogen receptor ␣, CT441-mediated SRAP1 degradation is not observed during the intracellular developmental cycle before host cells are lysed and infectious chlamydiae are released. Most compellingly, we have newly identified a chaperone activity of CT441, indicating a role of CT441 in prokaryotic protein quality control processes. Infections with the obligate intracellular pathogen Chlamydia trachomatis are among the most common sexually transmitted diseases worldwide, with approximately 1.5 million reported cases in the United States in 2012 (1). While most of the acute infections of the lower urogenital tract are asymptomatic and remain unrecognized by the affected people, ascending infections in females often result in severe chronic sequelae, such as pelvic inflammatory disease, ectopic pregnancy, and infertility (2). Despite its clinical relevance, many aspects of the underlying virulence mechanisms have not been elucidated so far.As for other pathogens, infectivity and the propensity to manipulate host immune responses largely depend on the repertoire of pathogenicity factors. The most extensively studied effector protein in Chlamydia research is CPAF (chlamydial protease-like activity factor), which has been reported to degrade a broad spectrum of host cell proteins (3). However, it has been shown that the observed degradation of many previously identified CPAF substrates is an artifact of the protein isolation process (4), and thus whether CPAF actively degrades host cell proteins during the intracellular developmental cycle is a controversial subject of discussion.A second chlamydial protease, designated CT441 for C. trachomatis, shares significant amino acid sequence similarity with tailspecific proteases (Tsps) from other species (e.g., 25% identity with Tsp from Escherichia coli) and was first proposed by Lad et al. to interfere with host antimicrobial and inflammatory responses (5, 6); however, in later reports, a role of CT441 and CPAF in the cleavage of NF-B during the chlamydial infection has been put into question (4, 7). Unique regions that show no similarities to any characterized domain are present at the N terminus of Tsp proteins. E. coli Tsp cleaves substrate proteins labeled with a Cterminal ssrA-encoded peptide tag (small stable RNA A) and is involved in ...
The prokaryotic obligate intracellular pathogen Chlamydia trachomatis is the most prevalent cause of preventable blindness, affecting approximately six million people worldwide. In addition, C. trachomatis is the most commonly reported sexually transmitted pathogen in Europe and the US, causing pelvic inflammation, ectopic pregnancy and infertility. As in other intracellular pathogens, proteases play crucial roles during most stages of the complex life cycle of Chlamydia. CT441 is a chlamydial protease that has been reported to interfere with oestrogen signalling of the host cell. Here, the recombinant production, purification and crystallization of an inactive variant of CT441, designated CT441 (active-site Ser455 replaced by Ala), are described. CT441 was crystallized in space group P22 1 2 1 , with unit-cell parameters a = 86.7, b = 184.0, c = 209.6 Å . A complete diffraction data set was collected to a resolution of 2.95 Å .
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