Genome-wide Analysis of Chlamydophila pneumoniae Gene Expression at the Late Stage of Infection

Miura, Kazumasa; Toh, Y.; Hirakawa, Hideki; Sugii, Manabu; Murata, Masayuki; Nakai, Kenta; Tashiro, Kosuke; Kuhara, Satoru; Azuma, Yoshinao; Shirai, Mutsunori

doi:10.1093/dnares/dsm032

Cited by 5 publications

(8 citation statements)

References 31 publications

(52 reference statements)

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“…Similar responses were observed using HeLa cells and MEFs (data not shown). In the absence of apoptotic stimuli, apoptotic induction by C. pneumoniae infection was not observed between 48 and 72 hpi, which are the middle and late stages of C. pneumoniae infection, respectively [ 40 ]; chlamydial infection partially stimulated STS-induced apoptosis at 72 hpi. These data, combined with previous results [ 18 – 21 ], indicate that C. pneumoniae infection at relatively low MOI represses STS-induced apoptosis of various host cell lines in the early-to-middle stages of infection, but not in the late stage.…”

Section: Resultsmentioning

confidence: 99%

“…To address how caspase-3 is not activated by infection-induced activated caspase-9, we first analyzed expression of genes encoding inhibitors of apoptosis protein IAP1, IAP2, and XIAP. DNA microarray and quantitative RT-PCR at 48 hpi (data not shown) [ 40 ], showed that expression of these genes was not increased. However, caspase-9 was co-localized with chlamydial inclusions at 48 hpi by immunofluorescence staining using a caspase-9 antibody with apaf - 1 −/− MEFs (Fig.…”

Section: Resultsmentioning

confidence: 99%

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An epistatic effect of apaf-1 and caspase-9 on chlamydial infection

et al. 2015

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Chlamydia is an obligate intracellular bacterial pathogen that replicates solely within a membrane-bound vacuole termed an inclusion. Chlamydia seems to perturb multiple cellular processes of the host, such as, rearrangement of the membrane trafficking system for its intracellular multiplication, and inhibition of host cell apoptosis for persistent infection. In an attempt to clarify host factor involvement in apoptosis regulation, we found that inhibition of Caspase-9 restricted, while Apaf-1 promoted, Chlamydia pneumoniae infection in HEp-2, HeLa, and mouse epithelial fibroblast (MEF) cells. These opposition contributions to the chlamydial infection were confirmed using caspase-9−/− and apaf-1−/− MEFs. Similar phenomena also appeared in the case of infection with Chlamydia trachomatis. Interestingly, caspase-9 in apaf-1−/− MEFs was activated by chlamydial infection but during the infection caspase-3 was not activated. That is, caspase-9 was activated without support for multiplication and activation by Apaf-1, and the activated caspase-9 may be physically disconnected from the caspase cascade. This may be partially explained by the observation of caspase-9 accumulation within chlamydial inclusions. The sequestration of caspase-9 by chlamydia seems to result in apoptosis repression, which is crucial for the chlamydial development cycle. Because Apaf-1 shares domains with intracellular innate immune receptor NOD1, it may play a key role in the strategy to regulate chlamydial infection.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

An epistatic effect of apaf-1 and caspase-9 on chlamydial infection

et al. 2015

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“…Miura et al . [ 16 ] searched for transcripts that are expressed at the late stage of the infection cycle and identified putative σ 28 promoters upstream of ltub and the annotated ORF CPn0332. According to the identified TSS, the putative σ 28 promoter postulated upstream of ORF CPn0332 is located inside the transcript.…”

Section: Resultsmentioning

confidence: 99%

“…Several σ 70 target genes such as ompA and omcB , could be verified experimentally [ 15 ]. The role of the two alternative σ factors is still unknown but some of the late genes expressed at the stage of RB-to-EB conversion seem to be directly regulated by σ 28 [ 16 - 18 ].…”

Section: Introductionmentioning

confidence: 99%

The transcriptional landscape of Chlamydia pneumoniae

et al. 2011

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BackgroundGene function analysis of the obligate intracellular bacterium Chlamydia pneumoniae is hampered by the facts that this organism is inaccessible to genetic manipulations and not cultivable outside the host. The genomes of several strains have been sequenced; however, very little information is available on the gene structure and transcriptome of C. pneumoniae.ResultsUsing a differential RNA-sequencing approach with specific enrichment of primary transcripts, we defined the transcriptome of purified elementary bodies and reticulate bodies of C. pneumoniae strain CWL-029; 565 transcriptional start sites of annotated genes and novel transcripts were mapped. Analysis of adjacent genes for co-transcription revealed 246 polycistronic transcripts. In total, a distinct transcription start site or an affiliation to an operon could be assigned to 862 out of 1,074 annotated protein coding genes. Semi-quantitative analysis of mapped cDNA reads revealed significant differences for 288 genes in the RNA levels of genes isolated from elementary bodies and reticulate bodies. We have identified and in part confirmed 75 novel putative non-coding RNAs. The detailed map of transcription start sites at single nucleotide resolution allowed for the first time a comprehensive and saturating analysis of promoter consensus sequences in Chlamydia.ConclusionsThe precise transcriptional landscape as a complement to the genome sequence will provide new insights into the organization, control and function of genes. Novel non-coding RNAs and identified common promoter motifs will help to understand gene regulation of this important human pathogen.

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“…To construct a chlamydial genome library for Y2H screening, DNA fragments coding chlamydial genes were individually amplified by PCR using C. pneumoniae J138 genomic DNA as a template (Miura et al, 2008) (Figs. 1 and 2a).…”

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confidence: 99%

Identification of <i>Chlamydia pneumoniae</i> candidate genes that interact with human apoptotic factor caspase-9

Aziz

Ushirokita

Azuma

2018

J. Gen. Appl. Microbiol.

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Chlamydia pneumoniae is an obligate intracellular pathogen responsible for respiratory diseases, including pneumonia and bronchitis, and is highly involved in chronic diseases, including atherosclerosis, asthma, and Alzheimer's disease. We previously showed that the host apoptotic factor caspase-9 played a crucial role for chlamydial multiplication and host apoptosis inhibition by chlamydial infection. To identify chlamydial genes interacting with human caspase-9, yeast two-hybrid screening was performed and 5 chlamydial genes, including Cpj0838 and pmpG were isolated from the C. pneumoniae genomic library. Pull-down experiments showed that caspase-9 physically bound to the Cpj0838 product and chlamydial cells, which contain PmpG proteins. This study could provide a clue to understanding host-Chlamydia interactions, especially the apoptosis repression by Chlamydia infection.

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Genome-wide Analysis of Chlamydophila pneumoniae Gene Expression at the Late Stage of Infection

Cited by 5 publications

References 31 publications

An epistatic effect of apaf-1 and caspase-9 on chlamydial infection

An epistatic effect of apaf-1 and caspase-9 on chlamydial infection

The transcriptional landscape of Chlamydia pneumoniae

Identification of <i>Chlamydia pneumoniae</i> candidate genes that interact with human apoptotic factor caspase-9

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