Identification of a serine protease inhibitor which causes inclusion vacuole reduction and is lethal to Chlamydia trachomatis

Published on Aug 1, 2013in Molecular Microbiology3.418
· DOI :10.1111/MMI.12306
Sarina Gloeckl4
Estimated H-index: 4
(QUT: Queensland University of Technology),
Vanissa A. Ong6
Estimated H-index: 6
(QUT: Queensland University of Technology)
+ 14 AuthorsWilhelmina M. Huston21
Estimated H-index: 21
(QUT: Queensland University of Technology)
The mechanistic details of the pathogenesis of Chlamydia, an obligate intracellular pathogen of global importance, have eluded scientists due to the scarcity of traditional molecular genetic tools to investigate this organism. Here we report a chemical biology strategy that has uncovered the first essential protease for this organism. Identification and application of a unique CtHtrA inhibitor (JO146) to cultures of Chlamydia resulted in a complete loss of viable elementary body formation. JO146 treatment during the replicative phase of development resulted in a loss of Chlamydia cell morphology, diminishing inclusion size, and ultimate loss of inclusions from the host cells. This completely prevented the formation of viable Chlamydia elementary bodies. In addition to its effect on the human Chlamydia trachomatis strain, JO146 inhibited the viability of the mouse strain, Chlamydia muridarum, both in vitro and in vivo. Thus, we report a chemical biology approach to establish an essential role for Chlamydia CtHtrA. The function of CtHtrA for Chlamydia appears to be essential for maintenance of cell morphology during replicative the phase and these findings provide proof of concept that proteases can be targeted for antimicrobial therapy for intracellular pathogens.
📖 Papers frequently viewed together
20107.50EMBO Reports
11 Authors (Benjamin Hoy, ..., Silja Wessler)
210 Citations
9 Authors (Manja Boehm, ..., Steffen Backert)
95 Citations
#1Michael P. Strauss (UTS: University of Technology, Sydney)H-Index: 2
#2Andrew T. F. LiewH-Index: 5
Last. Elizabeth J. Harry (UTS: University of Technology, Sydney)H-Index: 44
view all 6 authors...
FtsZ is a tubulin-like GTPase that is the major cytoskeletal protein in bacterial cell division. It polymerizes into a ring, called the Z ring, at the division site and acts as a scaffold to recruit other division proteins to this site as well as providing a contractile force for cytokinesis. To understand how FtsZ performs these functions, the in vivo architecture of the Z ring needs to be established, as well as how this structure constricts to enable cytokinesis. Conventional wide-field fluor...
181 CitationsSource
#1Allan L. Chen (UCI: University of California, Irvine)H-Index: 10
#2Kirsten A. Johnson (UCI: University of California, Irvine)H-Index: 5
Last. Ming Tan (UCI: University of California, Irvine)H-Index: 18
view all 5 authors...
Bacteria in the genus Chlamydia are major human pathogens that cause an intracellular infection. A chlamydial protease, CPAF, has been proposed as an important virulence factor that cleaves or degrades at least 16 host proteins, thereby altering multiple cellular processes. We examined 11 published CPAF substrates and found that there was no detectable proteolysis when CPAF activity was inhibited during cell processing. We show that the reported proteolysis of these putative CPAF substrates was ...
92 CitationsSource
#1Jan G. Christian (TUM: Technische Universität München)H-Index: 4
#2Julia Heymann (MPG: Max Planck Society)H-Index: 3
Last. Dagmar Heuer (MPG: Max Planck Society)H-Index: 15
view all 9 authors...
Chlamydiae are obligate intracellular bacteria that propagate in a cytosolic vacuole. Recent work has shown that growth of Chlamydia induces the fragmentation of the Golgi apparatus (GA) into ministacks, which facilitates the acquisition of host lipids into the growing inclusion. GA fragmentation results from infection-associated cleavage of the integral GA protein, golgin-84. Golgin-84-cleavage, GA fragmentation and growth of Chlamydia trachomatis can be blocked by the peptide inhibitor WEHD-fm...
41 CitationsSource
#1Yibing Wang (Southampton General Hospital)H-Index: 10
#2Simona Kahane (BGU: Ben-Gurion University of the Negev)H-Index: 19
Last. Ian N. Clarke (Southampton General Hospital)H-Index: 60
view all 6 authors...
Chlamydia trachomatis remains one of the few major human pathogens for which there is no transformation system. C. trachomatis has a unique obligate intracellular developmental cycle. The extracellular infectious elementary body (EB) is an infectious, electron-dense structure that, following host cell infection, differentiates into a non-infectious replicative form known as a reticulate body (RB). Host cells infected by C. trachomatis that are treated with penicillin are not lysed because this a...
281 CitationsSource
#1Wilhelmina M. Huston (QUT: Queensland University of Technology)H-Index: 21
#2Joel D. A. Tyndall (University of Otago)H-Index: 36
Last. Peter Timms (QUT: Queensland University of Technology)H-Index: 64
view all 5 authors...
DegP, a member of the HtrA family of proteins, conducts critical bacterial protein quality control by both chaperone and proteolysis activities. The regulatory mechanisms controlling these two distinct activities, however, are unknown. DegP activation is known to involve a unique mechanism of allosteric binding, conformational changes and oligomer formation. We have uncovered a novel role for the residues at the PDZ1:protease interface in oligomer formation specifically for chaperone substrates ...
25 CitationsSource
#1Theodor Chitlaru (Israel Institute for Biological Research)H-Index: 17
#2Galia Zaide (Israel Institute for Biological Research)H-Index: 4
Last. Avigdor Shafferman (Israel Institute for Biological Research)H-Index: 61
view all 6 authors...
Summary We demonstrate that disruption of the htrA (high temperature requirement A) gene in either the virulent Bacillus anthracis Vollum (pXO1+, pXO2+), or in the ΔVollum (pXO1-, pXO2-, nontoxinogenic and noncapsular) strains, affect significantly the ability of the resulting mutants to withstand heat, oxidative, ethanol and osmotic stress. The ΔhtrA mutants manifest altered secretion of several proteins, as well as complete silencing of the abundant extracellular starvation-associated neutral ...
43 CitationsSource
#1Carolyn I. Hall (Stanford University)H-Index: 2
#2Michael L. Reese (Stanford University)H-Index: 17
Last. Matthew Bogyo (Stanford University)H-Index: 88
view all 13 authors...
Toxoplasma gondii is a member of the phylum Apicomplexa that includes several important human pathogens, such as Cryptosporidium and Plasmodium falciparum, the causative agent of human malaria. It is an obligate intracellular parasite that can cause severe disease in congenitally infected neonates and immunocompromised individuals. Despite the importance of attachment and invasion to the success of the parasite, little is known about the underlying mechanisms that drive these processes. Here we ...
53 CitationsSource
#1Xiang Wu (University of Texas Health Science Center at San Antonio)H-Index: 1
#2Lei Lei (University of Texas Health Science Center at San Antonio)H-Index: 19
Last. Guangming Zhong (University of Texas Health Science Center at San Antonio)H-Index: 66
view all 6 authors...
The periplasmic High Temperature Requirement protein A (HtrA) plays important roles in bacterial protein folding and stress responses. However, the role of chlamydial HtrA (cHtrA) in chlamydial pathogenesis is not clear. The cHtrA was detected both inside and outside the chlamydial inclusions. The detection was specific since both polyclonal and monoclonal anti-cHtrA antibodies revealed similar intracellular labeling patterns that were only removed by absorption with cHtrA but not control fusion...
49 CitationsSource
#1Laszlo Kari (NIH: National Institutes of Health)H-Index: 21
#2Morgan M. GoheenH-Index: 7
Last. Harlan D. CaldwellH-Index: 63
view all 12 authors...
Chlamydia trachomatis is an obligate intracellular bacterial pathogen that infects hundreds of millions of individuals globally, causing blinding trachoma and sexually transmitted disease. More effective chlamydial control measures are needed, but progress toward this end has been severely hampered by the lack of a tenable chlamydial genetic system. Here, we describe a reverse-genetic approach to create isogenic C. trachomatis mutants. C. trachomatis was subjected to low-level ethyl methanesulfo...
127 CitationsSource
#1Tim Clausen (IMP: Research Institute of Molecular Pathology)H-Index: 53
#2Markus KaiserH-Index: 46
Last. Michael Ehrmann (Cardiff University)H-Index: 51
view all 4 authors...
Controlled proteolysis underlies a vast diversity of protective and regulatory processes that are of key importance to cell fate. The unique molecular architecture of the widely conserved high temperature requirement A (HTRA) proteases has evolved to mediate critical aspects of ATP-independent protein quality control. The simple combination of a classic Ser protease domain and a carboxy-terminal peptide-binding domain produces cellular factors of remarkable structural and functional plasticity t...
334 CitationsSource
Cited By40
#1Jimin Hwang (University of Otago)H-Index: 1
#2Natalie Strange (UTS: University of Technology, Sydney)H-Index: 2
Last. Joel D. A. Tyndall (University of Otago)H-Index: 36
view all 8 authors...
Abstract null null The obligate intracellular bacterium Chlamydia trachomatis (C. trachomatis) is responsible for the most common bacterial sexually transmitted infection and is the leading cause of preventable blindness, representing a major global health burden. While C. trachomatis infection is currently treatable with broad-spectrum antibiotics, there would be many benefits of a chlamydia-specific therapy. Previously, we have identified a small-molecule lead compound JO146 [Boc-Val-Pro-ValP(...
#1Karissa J. Munoz (UCI: University of California, Irvine)H-Index: 1
#2Kevin Wang (UCI: University of California, Irvine)H-Index: 1
Last. Christine Sütterlin (UCI: University of California, Irvine)H-Index: 22
view all 5 authors...
Chlamydia is an obligate intracellular bacterium and the most common reportable cause of human infection in the U.S. This pathogen proliferates inside a eukaryotic host cell, where it resides within a membrane-bound compartment called the chlamydial inclusion. It has an unusual developmental cycle, marked by conversion between a replicating form, the reticulate body (RB), and an infectious form, the elementary body (EB). We found that the small molecule H89 slowed inclusion growth and decreased ...
#1Ruo-Yi Xue (Third Military Medical University)H-Index: 2
#2Chang Liu (Third Military Medical University)H-Index: 2
Last. Li Haibo
view all 6 authors...
Abstract Background As there is an urgent need to develop antibacterial therapies other than antibiotics, more and more researches have focused on the high temperature requirement protein A (HtrA) family proteases, which have both serine protease and chaperone activities. Objectives The research progresses of the role of HtrA family proteases in the pathogenesis of bacterial infections are summarized, and the "pros and cons" of exploiting HtrA inhibitors in antibacterial drug development are pro...
1 CitationsSource
#1Brenda B. Daroz (Instituto Butantan)H-Index: 1
#2Luis G. V. Fernandes (Instituto Butantan)H-Index: 11
Last. Ana L. T. O. Nascimento (Instituto Butantan)H-Index: 34
view all 4 authors...
Leptospirosis is a zoonosis caused by the pathogenic bacteria of the genus Leptospira. The identification of conserved outer membrane proteins among pathogenic strains is a major research target in elucidating mechanisms of pathogenicity. Surface-exposed proteins are most probably the ones involved in the interaction of leptospires with the environment. Some spirochetes use outer membrane proteases as a way to penetrate host tissues. HtrA is a family of proteins found in various cell types, from...
1 CitationsSource
#1Xiaohua Dong (HBNU: Hebei North University)H-Index: 1
#2Wanxing Zhang (Tianjin Medical University General Hospital)H-Index: 1
Last. Shuping Hou (Tianjin Medical University General Hospital)H-Index: 2
view all 7 authors...
BACKGROUND Chlamydia trachomatis is an obligate intracellular pathogen that can cause severe reproductive tract complications while ascending infection occurs. When spreading from cell to cell in a host, C. trachomatis utilizes various survival strategies to offset host defense mechanisms. One such strategy is to degrade host antimicrobial defense proteins before they can attack the invading C. trachomatis cells. MATERIAL AND METHODS We expressed and purified recombinant chlamydia high temperatu...
3 CitationsSource
#1Ayodeji A. Agbowuro (University of Otago)H-Index: 2
#2Jimin Hwang (University of Otago)H-Index: 1
Last. Joel D. A. Tyndall (University of Otago)H-Index: 36
view all 10 authors...
Abstract Chlamydia trachomatis high temperature requirement A (CtHtrA) is a serine protease that performs proteolytic and chaperone functions in pathogenic Chlamydiae; and is seen as a prospective drug target. This study details the strategies employed in optimizing the irreversible CtHtrA inhibitor JO146 [Boc-Val-Pro-ValP(OPh)2] for potency and selectivity. A series of adaptations both at the warhead and specificity residues P1 and P3 yielded 23 analogues, which were tested in human neutrophil ...
2 CitationsSource
#1Yachun Zhou (CSU: Central South University)H-Index: 1
#2Xiaofang Lu (CSU: Central South University)H-Index: 2
Last. Yong Wang (CSU: Central South University)H-Index: 4
view all 9 authors...
Abstract Chlamydia (C.) trachomatis, characterized by a unique biphasic life cycle, is an obligate intracellular bacterial pathogen which is responsible for the highest number of sexually transmitted bacterial infections globally. However, its pathogenic mechanisms have not been fully elucidated because of its unique developmental cycle and obligate intracellular nature. High temperature requirement (HtrA), a critical protease and chaperone, has been previously demonstrated to be essential for s...
1 CitationsSource
#1Gerardo García-González (Universidad Autónoma de Nuevo León)H-Index: 1
#2Gloria M. González (Universidad Autónoma de Nuevo León)H-Index: 28
Last. José Prisco Palma-Nicolás (Universidad Autónoma de Nuevo León)H-Index: 10
view all 3 authors...
Resumen Las proteasas de serina son enzimas ampliamente distribuidas en la naturaleza, responsables de multiples e importantes procesos biologicos. Durante las infecciones bacterianas los patogenos secretan y usan sus proteasas de serina como factores de virulencia para combatir contra el huesped, a traves de diversos efectos como la desorganizacion de tejidos, la proteolisis de efectores inmunologicos o la inactivacion de componentes relevantes para la fisiologia del huesped; sin embargo, desde...
: Infection with Helicobacter pylori represents a major risk for developing peptic ulcer disease, gastric adenocarcinoma, and various other gastric and nongastric sicknesses. A series of H. pylori virulence factors can be secreted into the cell culture supernatant, and the secretome contains more than 100 different proteins. However, the quantities of proteins secreted by the bacteria over time are unknown. One of these factors is the serine protease high-temperature requirement A (HtrA), encode...
4 CitationsSource
#1Regan J. Hayward (UTS: University of Technology, Sydney)H-Index: 4
#2James W. Marsh (MPG: Max Planck Society)H-Index: 3
Last. Garry S. A. Myers (UTS: University of Technology, Sydney)H-Index: 31
view all 5 authors...
Chlamydia are Gram-negative, obligate intracellular bacterial pathogens responsible for a broad spectrum of human and animal diseases. In humans, Chlamydia trachomatis is the most prevalent bacterial sexually transmitted infection worldwide and is the causative agent of trachoma (infectious blindness) in disadvantaged populations. Over the course of its developmental cycle, Chlamydia extensively remodels its intracellular niche and parasitises the host cell for nutrients, with substantial result...
2 CitationsSource