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pmid:40444155

The Porphyromonas gingivalis RNA-binding protein is required for growth in high levels of zinc and persistence with host cells.

Sai Yanamandra · Holly Marsh · Romana Cvitkovic · Qin Gui · Benjamin R Belvin · Janina P Lewis

Frontiers in cellular and infection microbiology · 2025 doi:10.3389/fcimb.2025.1569544

Paper

complete

Experiment

complete

Data

partial

Analysis

partial

Execution

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title
The Porphyromonas gingivalis RNA-binding protein is required for growth in high levels of zinc and persistence with host cells.
doi
10.3389/fcimb.2025.1569544
pmid
40444155
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[0]
Sai Yanamandra
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Holly Marsh
[2]
Romana Cvitkovic
[3]
Qin Gui
[4]
Benjamin R Belvin
[5]
Janina P Lewis
journal
Frontiers in cellular and infection microbiology
year
2025
abstract
The oral periodontal pathogen Porphyromonas gingivalis must adapt to an ever-changing environment to survive and cause disease. So far, most of the efforts concerning the regulatory mechanisms employed by the bacterium centered on DNA-binding regulators. Although global regulatory mechanisms employing RNA-binding proteins (RBP) are reported in most forms of life so far, such mechanism of regulation remains unknown in the oral Bacteroidetes group. Examination of the genome of P. gingivalis led to the discovery of a putative RBP with the RNA recognition motif 1 (RRM-1) designated here RbpPg1 (RNA-binding protein Porphyromonas gingivalis 1). The recombinant form of the protein-bound RNA and RNA-pull down identified a zinc exporter transcript as the most enriched one in agreement with the higher levels of zinc in the absence of the protein. Deletion of RbpPg1 reduced the ability of the bacterium to grow with 0.5 mM zinc. The RgpB protein level and the Arg-X protease activity was reduced in both iron replete and iron deplete conditions in the mutant strain when compared to the wild type. Lys-X protease activity was reduced, although Kgp protein levels were not altered by deletion of RbpPg1. The mutant grew better in hemin-deplete conditions when compared to the wild type. Finally, RbpPg1 was indispensable for the bacterium to survive with host cells. We have determined that both the transcriptome and proteome are affected by the deletion of RbpPg1 and found that the major group of proteins with elevated expression were the ones associated with response to environmental stress changes, while proteins mediating metabolic processes were downregulated. Overall, the first RBP characterized in P. gingivalis plays a significant role in the biology of the bacterium and differs from RBPs in other Gram-negative bacteria. Data are available via ProteomeXchange with identifier PXD034144 and via the NCBI Gene Expression Omnibus (GEO) and under accession number GSE168570.
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p-Value was calculated using Student’s t-test. q-Value was calculated using significance analysis of microarrays (SAM) (Tusher et al., 2001) and permutation-based false discovery rate (FDR).
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The RNAseq data have been deposited to the NCBI Gene Expression Omnibus (GEO) and available under the accession number GSE168570 (“Role of RNAbindingprotein, Pgr, in P. gingivalis”). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partnerrepository with the dataset identifier PXD034144 (“Role of RNA binding protein PgRBP1 in alteration of protein profile of Porphyromonas gingivalis”).
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The RNAseq data have been deposited to the NCBI Gene Expression Omnibus (GEO) and available under the accession number GSE168570 (“Role of RNAbindingprotein, Pgr, in P. gingivalis”). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partnerrepository with the dataset identifier PXD034144 (“Role of RNA binding protein PgRBP1 in alteration of protein profile of Porphyromonas gingivalis”).
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We observed a 10-fold reduction in survival of the RbpPg1-deficient strain in the presence of host cells (survival of both attached and internalized bacteria) when compared to the parental W83 bacteria ( Figure 7A ).
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Finally, RbpPg1 was indispensable for the bacterium to survive with host cells.
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We observed a 10-fold reduction in survival of the RbpPg1-deficient strain in the presence of host cells (survival of both attached and internalized bacteria) when compared to the parental W83 bacteria ( Figure 7A ).
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The RNAseq data have been deposited to the NCBI Gene Expression Omnibus (GEO) and available under the accession number GSE168570 (“Role of RNAbindingprotein, Pgr, in P. gingivalis”). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partnerrepository with the dataset identifier PXD034144 (“Role of RNA binding protein PgRBP1 in alteration of protein profile of Porphyromonas gingivalis”).
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Role of RNA-binding protein, Pgr, in Porphyromonas gingivalis
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The RNAseq data have been deposited to the NCBI Gene Expression Omnibus (GEO) and available under the accession number GSE168570 (“Role of RNAbindingprotein, Pgr, in P. gingivalis”). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partnerrepository with the dataset identifier PXD034144 (“Role of RNA binding protein PgRBP1 in alteration of protein profile of Porphyromonas gingivalis”).
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Role of RNA binding protein PgRBP1 in alteration of protein profile of Porphyromonas gingivalis
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Complete file
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So far, most of the efforts concerning the regulatory mechanisms employed by the bacterium centered on DNA-binding regulators. Although global regulatory mechanisms employing RNA-binding proteins (RBP) are reported in most forms of life so far, such mechanism of regulation remains unknown in the oral Bacteroidetes group. Examination of the genome of P. gingivalis led to the discovery of a putative RBP with the RNA recognition motif 1 (RRM-1) designated here RbpPg1 (RNA-binding protein Porphyromonas gingivalis 1). The recombinant form of the protein-bound RNA and RNA-pull down identified a zinc exporter transcript as the most enriched one in agreement with the higher levels of zinc in the absence of the protein. Deletion of RbpPg1 reduced the ability of the bacterium to grow with 0.5 mM zinc. The RgpB protein level and the Arg-X protease activity was reduced in both iron replete and iron deplete conditions in the mutant strain when compared to the wild type. Lys-X protease activity was reduced, although Kgp protein levels were not altered by deletion of RbpPg1. The mutant grew better in hemin-deplete conditions when compared to the wild type. Finally, RbpPg1 was indispensable for the bacterium to survive with host cells. We have determined that both the transcriptome and proteome are affected by the deletion of RbpPg1 and found that the major group of proteins with elevated expression were the ones associated with response to environmental stress changes, while proteins mediating metabolic processes were downregulated. Overall, the first RBP characterized in P. gingivalis plays a significant role in the biology of the bacterium and differs from RBPs in other Gram-negative bacteria. 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