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PMID:25158041
| Citation |
Hare, RF and Hueffer, K (2014) Francisella novicida pathogenicity island encoded proteins were secreted during infection of macrophage-like cells. PLoS ONE 9:e105773 |
|---|---|
| Abstract |
Intracellular pathogens and other organisms have evolved mechanisms to exploit host cells for their life cycles. Virulence genes of some intracellular bacteria responsible for these mechanisms are located in pathogenicity islands, such as secretion systems that secrete effector proteins. The Francisella pathogenicity island is required for phagosomal escape, intracellular replication, evasion of host immune responses, virulence, and encodes a type 6 secretion system. We hypothesize that some Francisella novicida pathogenicity island proteins are secreted during infection of host cells. To test this hypothesis, expression plasmids for all Francisella novicida FPI-encoded proteins with C-terminal and N-terminal epitope FLAG tags were developed. These plasmids expressed their respective epitope FLAG-tagged proteins at their predicted molecular weights. J774 murine macrophage-like cells were infected with Francisella novicida containing these plasmids. The FPI proteins expressed from these plasmids successfully restored the intramacrophage growth phenotype in mutants of the respective genes that were deficient for intramacrophage growth. Using these expression plasmids, the localization of the Francisella pathogenicity island proteins were examined via immuno-fluorescence microscopy within infected macrophage-like cells. Several Francisella pathogenicity island encoded proteins (IglABCDEFGHIJ, PdpACE, DotU and VgrG) were detected extracellularly and they were co-localized with the bacteria, while PdpBD and Anmk were not detected and thus remained inside bacteria. Proteins that were co-localized with bacteria had different patterns of localization. The localization of IglC was dependent on the type 6 secretion system. This suggests that some Francisella pathogenicity island proteins were secreted while others remain within the bacterium during infection of host cells as structural components of the secretion system and were necessary for secretion. |
| Links |
PubMed PMC4144950 Online version:10.1371/journal.pone.0105773 |
| Keywords |
Animals; Bacterial Proteins/genetics; Bacterial Proteins/secretion; Cell Line; Francisella/genetics; Francisella/pathogenicity; Genes, Bacterial; Genomic Islands; Macrophages/microbiology; Mice; Protein Transport; Type VI Secretion Systems/metabolism; Virulence Factors/genetics |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages "[IglE]... were co-localized with bacteria and also extending beyond, completely surrounding bacteria." |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages "[IglE]... were co-localized with bacteria and also extending beyond, completely surrounding bacteria." |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (top right panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case IglE is the protein being addressed. |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages “IglA was localized with bacteria.” |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (5th row left panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case IglA is the protein being addressed. |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages “...PdpA [was] also localized with bacteria, and on occasion surrounding the bacteria." |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages “...PdpA were also localized with bacteria, and on occasion surrounding the bacteria." |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (top left panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case PdpA is the protein being adressed. |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages "[IglC] were co-localized with bacteria and also extending beyond, completely surrounding bacteria." |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages "[IglC]... [was] co-localized with bacteria and also extending beyond, completely surrounding bacteria." |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (4th row middle panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case IglC is the protein being addressed. |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages “IglD... were also localized with bacteria, and on occasion surrounding the bacteria." |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages “IglD… [was] also localized with bacteria, and on occasion surrounding the bacteria." |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (4th row left panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case IglD is the protein being addressed. |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages Looking at the figure it is shown that the protein IglB is localized with the bacteria |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (4th row right panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case IglB is the protein being addressed. |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Not shown in figure but implied by text. "PdpC was also detected both co-localized with bacteria and dispersing away from the bacteria." |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages Looking at the figure it is shown that the protein IglI is localized with the bacteria. |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages Looking at the figure it is shown that the protein IglI is surrounding the bacteria. |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (3rd row middle panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case IglI is the protein being addressed. |
complete | ||||
| GO:0030430: host cell cytoplasm |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages "VgrG were co-localized with bacteria and also extending beyond, completely surrounding bacteria." |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 3: Localization of FPI Proteins within Infected Macrophages "VgrG [was] co-localized with bacteria and also extending beyond, completely surrounding bacteria." |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (2nd row left panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case VgrG is the protein being addressed. |
complete | ||||
| GO:0106139: symbiont cell surface |
ECO:0000314: |
C |
Organism is Francisella novicida. Figure 4 Localization of FPI Proteins During Infection of Sua-1B Cells. "The C-FLAG PdpE was studded around the bacterium..." |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (top middle panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case PdpB is the protein being addressed. |
complete | ||||
| GO:0061887: reproduction of symbiont in host |
ECO:0000315: |
P |
The organism is Francisella novicida. Figure 2 (3rd row left panel) "The FPI mutant lacks all FPI genes and was defective for intracellular growth, as were each of the individual FPI gene mutants." In this case DotU is the protein being addressed. |
complete | ||||
Notes
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