Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells

ABSTRACT A defining characteristic of Chlamydia spp. is their developmental cycle characterized by outer membrane transformations of cysteine bonds among cysteine-rich outer membrane proteins. The reduction-oxidation states of host cell compartments were monitored during the developmental cycle usin...

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Autores principales:	Xiaogang Wang, Christian Schwarzer, Kevin Hybiske, Terry E. Machen, Richard S. Stephens
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2014
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/2b52c1422e2c4a909e862af927a06c15
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spelling	oai:doaj.org-article:2b52c1422e2c4a909e862af927a06c152021-11-15T15:47:03ZDevelopmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells10.1128/mBio.01924-142150-7511https://doaj.org/article/2b52c1422e2c4a909e862af927a06c152014-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01924-14https://doaj.org/toc/2150-7511ABSTRACT A defining characteristic of Chlamydia spp. is their developmental cycle characterized by outer membrane transformations of cysteine bonds among cysteine-rich outer membrane proteins. The reduction-oxidation states of host cell compartments were monitored during the developmental cycle using live fluorescence microscopy. Organelle redox states were studied using redox-sensitive green fluorescent protein (roGFP1) expressed in CF15 epithelial cells and targeted to the cytosol, mitochondria, and endoplasmic reticulum (ER). The redox properties of chlamydiae and the inclusion were monitored using roGFP expressed by Chlamydia trachomatis following transformation. Despite the large morphological changes associated with chlamydial infection, redox potentials of the cytosol (Ψcyto [average, −320 mV]), mitochondria (Ψmito [average, −345 mV]), and the ER (ΨER [average, −258 mV]) and their characteristic redox regulatory abilities remained unchanged until the cells died, at which point Ψcyto and Ψmito became more oxidized and ΨER became more reduced. The redox status of the chamydial cytoplasm was measured following transformation and expression of the roGFP biosensor in C. trachomatis throughout the developmental cycle. The periplasmic and outer membrane redox states were assessed by the level of cysteine cross-linking of cysteine-rich envelope proteins. In both cases, the chlamydiae were highly reduced early in the developmental cycle and became oxidized late in the developmental cycle. The production of a late-developmental-stage oxidoreductase/isomerase, DsbJ, may play a key role in the regulation of the oxidoreductive developmental-stage-specific process. IMPORTANCE Infectious Chlamydia organisms have highly oxidized and cysteine cross-linked membrane proteins that confer environmental stability when outside their host cells. Once these organisms infect a new host cell, the proteins become reduced and remain reduced during the active growth stage. These proteins become oxidized at the end of their growth cycle, wherein infectious organisms are produced and released to the environment. How chlamydiae mediate and regulate this key step in their pathogenesis is unknown. Using biosensors specifically targeted to different compartments within the infected host cell and for the chlamydial organisms themselves, the oxidoreductive states of these compartments were measured during the course of infection. We found that the host cell redox states are not changed by infection with C. trachomatis, whereas the state of the chlamydial organisms remains reduced during infection until the late developmental stages, wherein the organisms’ cytosol and periplasm become oxidized and they acquire environmental resistance and infectivity.Xiaogang WangChristian SchwarzerKevin HybiskeTerry E. MachenRichard S. StephensAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 6 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Xiaogang Wang Christian Schwarzer Kevin Hybiske Terry E. Machen Richard S. Stephens Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells
description	ABSTRACT A defining characteristic of Chlamydia spp. is their developmental cycle characterized by outer membrane transformations of cysteine bonds among cysteine-rich outer membrane proteins. The reduction-oxidation states of host cell compartments were monitored during the developmental cycle using live fluorescence microscopy. Organelle redox states were studied using redox-sensitive green fluorescent protein (roGFP1) expressed in CF15 epithelial cells and targeted to the cytosol, mitochondria, and endoplasmic reticulum (ER). The redox properties of chlamydiae and the inclusion were monitored using roGFP expressed by Chlamydia trachomatis following transformation. Despite the large morphological changes associated with chlamydial infection, redox potentials of the cytosol (Ψcyto [average, −320 mV]), mitochondria (Ψmito [average, −345 mV]), and the ER (ΨER [average, −258 mV]) and their characteristic redox regulatory abilities remained unchanged until the cells died, at which point Ψcyto and Ψmito became more oxidized and ΨER became more reduced. The redox status of the chamydial cytoplasm was measured following transformation and expression of the roGFP biosensor in C. trachomatis throughout the developmental cycle. The periplasmic and outer membrane redox states were assessed by the level of cysteine cross-linking of cysteine-rich envelope proteins. In both cases, the chlamydiae were highly reduced early in the developmental cycle and became oxidized late in the developmental cycle. The production of a late-developmental-stage oxidoreductase/isomerase, DsbJ, may play a key role in the regulation of the oxidoreductive developmental-stage-specific process. IMPORTANCE Infectious Chlamydia organisms have highly oxidized and cysteine cross-linked membrane proteins that confer environmental stability when outside their host cells. Once these organisms infect a new host cell, the proteins become reduced and remain reduced during the active growth stage. These proteins become oxidized at the end of their growth cycle, wherein infectious organisms are produced and released to the environment. How chlamydiae mediate and regulate this key step in their pathogenesis is unknown. Using biosensors specifically targeted to different compartments within the infected host cell and for the chlamydial organisms themselves, the oxidoreductive states of these compartments were measured during the course of infection. We found that the host cell redox states are not changed by infection with C. trachomatis, whereas the state of the chlamydial organisms remains reduced during infection until the late developmental stages, wherein the organisms’ cytosol and periplasm become oxidized and they acquire environmental resistance and infectivity.
format	article
author	Xiaogang Wang Christian Schwarzer Kevin Hybiske Terry E. Machen Richard S. Stephens
author_facet	Xiaogang Wang Christian Schwarzer Kevin Hybiske Terry E. Machen Richard S. Stephens
author_sort	Xiaogang Wang
title	Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells
title_short	Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells
title_full	Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells
title_fullStr	Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells
title_full_unstemmed	Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells
title_sort	developmental stage oxidoreductive states of <italic toggle="yes">chlamydia</italic> and infected host cells
publisher	American Society for Microbiology
publishDate	2014
url	https://doaj.org/article/2b52c1422e2c4a909e862af927a06c15
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Developmental Stage Oxidoreductive States of <italic toggle="yes">Chlamydia</italic> and Infected Host Cells

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