Evolutionarily conserved regulation of immunity by the splicing factor RNP-6/PUF60
- Max Planck Institute for Biology of Ageing, Germany
- Department I of Internal Medicine, University of Cologne, Germany
- Division of Infectious Diseases, University of Cologne, Germany
- German Center for Infection Research (DZIF), Partner Site Bonn-Cologne, Germany
- Center for Molecular Medicine Cologne (CMMC), Germany
- Cellular-Stress and Immune Response Laboratory, Centre for Cancer Biology, University of South Australia, Australia
- Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Germany
Figures
Figure 1 with 1 supplement
Isolation of the rnp-6 G281D substitution mutant from a cold resistance screen.
(A) Cold stress survival assay. Low temperature (2°C) incubation kills wildtype young adult worms. (B) daf-2 and eat-2 mutants show enhanced survival after 24 hr incubation at 2°C. (C) Schematic of …
Figure 1—figure supplement 1
rnp-6 G281D mutation compromises bacterial infection survival, related to Figure 1.
(A) Number of progenies per animal of wildtype and rnp-6(dh1127) worms. rnp-6(dh1127) mutants have a smaller brood size. (B) Development time of wildtype and rnp-6(dh1127) animals. rnp-6(dh1127) …
Figure 2 with 1 supplement
Dysregulation of immune responses in the rnp-6 G281D mutant.
(A,B) GO enrichment analysis using DAVID. RNA sequencing was performed on WT and rnp-6(dh1127) animals under control and S. aureus infected conditions. The 10 most statistically significant terms …
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Figure 2—source data 1
List of differentially expressed genes.
Non-infected.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig2-data1-v2.xlsx
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Figure 2—source data 2
List of differentially expressed genes.
Infected.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig2-data2-v2.xlsx
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Figure 2—source data 3
List of rnp-6 dependent infection induced genes.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig2-data3-v2.xlsx
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Figure 2—source data 4
List of rnp-6 dependent infection repressed genes.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig2-data4-v2.xlsx
Figure 2—figure supplement 1
G281D substitution of rnp-6 inhibits transcriptional responses to bacterial infection, related to Figure 2.
(A,B) GO enrichment analysis of rnp-6 dependent infection responsive genes. (C) qRT-PCR results showing rnp-6 G281D substitution compromises induction of infection responsive genes upon S. aureus …
Figure 3 with 1 supplement
rnp-6 remodels alternate splicing patterns upon infection.
(A) Venn diagrams showing the numbers of alternatively spliced genes induced by S. aureus infection and those that are affected in infected rnp-6(dh1127) when compared to infected N2 animals. (B,C,D)…
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Figure 3—source data 1
List of alternatively spliced genes upon S. aureus infection in wildtype animals.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig3-data1-v2.xlsx
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Figure 3—source data 2
List of alternatively spliced genes between infected wildtype animals and infected rnp-6(dh1127) mutants.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig3-data2-v2.xlsx
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Figure 3—source data 3
List of genes whose splicing is affected by both infection and rnp-6.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig3-data3-v2.xlsx
Figure 3—figure supplement 1
Alternative splicing events induced by infection and rnp-6 G281D substitution, related to Figure 3.
(A,B) Pie charts showing the percentage of each type of alternative splicing events in each indicated comparison. Analysis was performed with SAJR method. The numbers of splicing events are depicted.
Figure 4 with 1 supplement
RNP-6 restrains innate immunity in C. elegans.
(A) Infection survival analysis. Young adult worms fed with either control or rnp-6 RNAi HT115 bacteria were infected with S. aureus. rnp-6 RNAi leads to prolonged survival (p<0.0001, log-rank …
Figure 4—figure supplement 1
rnp-6 regulates immune gene expression, and its overexpression compromises infection survival, related to Figure 4.
(A) Infection survival analysis. rnp-6 OP50 RNAi prolongs survival of N2 animals under full lawn S. aureus infection condition (p<0.0001, log-rank test.). (B) Bacterial burden at 24 hr …
Figure 5 with 2 supplements
RNP-6 controls PMK-1 activity.
(A) Pie charts showing the portion of rnp-6 dependent infection responsive genes which are also reported to be pmk-1 dependent. 17 out of 116 rnp-6 dependent infection induced genes also have pmk-1 …
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Figure 5—source data 1
List of rnp-6 dependent infection induced genes with pmk-1 dependency.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig5-data1-v2.xlsx
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Figure 5—source data 2
List of rnp-6 dependent infection repressed genes with pmk-1 dependency.
- https://cdn.elifesciences.org/articles/57591/elife-57591-fig5-data2-v2.xlsx
Figure 5—figure supplement 1
daf-16 and hlh-30 do not play a significant role in rnp-6 mediated immunity, related to Figure 5.
(A,B) Infection survival analysis. rnp-6 OP50 RNAi prolongs survival under S. aureus infection condition in daf-16 (p<0.0001) and hlh-30 (p<0.0001, log-rank test.) mutants.
Figure 5—figure supplement 2
rnp-6 regulates immunity independently of splicing of the PMK-1 MAPK signaling pathway genes, related to Figure 5.
(A–I) Sequencing reads coverage for gene members of the PMK-1 MAPK signaling pathway. These genes are tir-1, nsy-1, sek-1, pmk-1, vhp-1, skn-1, atf-7, riok-1 and cebp-1. The splicing patterns of …
Figure 6 with 1 supplement
Suppression of immunity by SFA-1.
(A) Demographic analysis of lifespan. Overexpression of SFA-1 extends lifespan when the animals are cultured withE. coliOP50 at 25°C (p<0.0001, log-rank test.). (B) Infection survival analysis. …
Figure 6—figure supplement 1
sfa-1 specifically work with rnp-6 to regulate infection survival, related to Figure 6.
(A) Infection survival analysis. SFA-1 overexpression causes sensitivity to E. faecalis (p<0.0001, log-rank test.). (B,C) repo-1 or hrpu-1 RNAi does not fully rescue the sensitivity of rnp-6(dh1127) …
Figure 7 with 1 supplement
Evolutionary conservation of immuno-suppressive and anti-inflammatory functions of PUF60 in mammalian cells.
(A) Western blot analysis of S. aureus infected HeLa, MEF and RAW264.7 cells at 6 hr post-infection. S. aureus infection significantly reduces the protein levels of PUF60. (B) qRT-PCR results. HeLa …
Figure 7—figure supplement 1
PUF60 suppresses inflammation in mammalian cells, related to Figure 7.
(A) qRT-PCR results of S. aureus infected HeLa cells. At post-infection 6 hr, PUF60 transcript levels are significantly lower in the infected cells. (B) Western blot showing PUF60 levels are reduced …
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Additional files
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Supplementary file 1
List of primers.
- https://cdn.elifesciences.org/articles/57591/elife-57591-supp1-v2.docx
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Supplementary file 2
qPCR primer binding sites for C. elegans targets.
- https://cdn.elifesciences.org/articles/57591/elife-57591-supp2-v2.docx
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Supplementary file 3
qPCR primer binding sites for human targets.
- https://cdn.elifesciences.org/articles/57591/elife-57591-supp3-v2.docx
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Supplementary file 4
qPCR primer binding sites for mouse targets.
- https://cdn.elifesciences.org/articles/57591/elife-57591-supp4-v2.docx
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Supplementary file 5
List of antibodies.
- https://cdn.elifesciences.org/articles/57591/elife-57591-supp5-v2.docx
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Supplementary file 6
Table of survival data.
- https://cdn.elifesciences.org/articles/57591/elife-57591-supp6-v2.docx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/57591/elife-57591-transrepform-v2.docx
