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Abstract
Nuclear targeting of bacterial proteins in host cells and subsequent interaction with nuclear molecules are an emerging pathogenic mechanism of bacteria. In this study, we predicted the nuclear targeting proteins with nuclear localization signals (NLSs) in Staphylococcus aureus using bioinformatic analysis. A total of 51 proteins of S. aureus, comprising of 24 functional and 27 hypothetical proteins, were predicted to carry putative NLSs. Among them, β-lactamase and MsrR proteins with the putative NLSs were selected to determine the nuclear targeting in host cells. Fusion proteins of BlaZ-green fluorescent protein (GFP) were evenly distributed in the nuclei of host cells and subsequently induced host cell death. However, fusion proteins of MsrR-GFP were not localized in the nuclei of host cells In conclusion, screening of nuclear targeting proteins with NLSs and determination of their pathology in host cells may open up the new field of S. aureus pathogenesis.
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Figure 1.
Nuclear targeting of fusion proteins of BlaZ-GFP in host cells. COS-7 and HEp-2 cells were transfected with blaZ and msrR cloned with pAcGFP1-N2 vector, respectively. Cells were transfected with the empty pAcGFP1-N2 vector as a control. Expression of BlaZ-GFP fusion proteins in host cells was observed using fluorescence microscope. Magnification: ×40.
Figure 2.
Cytosolic localization of fusion proteins of GFP-BlaZ in host cells. COS-7 cells were transfected with pAcGFP1-C2 vector or blaZ cloned with pAcGFP1-C2 vector. Expression of GFP-BlaZ fusion proteins in host cells was observed using fluorescence microscope. Magnification: ×40.
Figure 3.
Cytotoxicity of β-lactamase fused with GFP in host cells. Cells were transfected with plasmid constructs of blaZ cloned with pAcGFP1-C2 and pAcGFP1-N2 vector, respectively. Cells were transfected with pAcGFP1-N2 or pAcGFP1-C2 vectors as a control. Cells transfected cells with plasmid constructs were incubated at 37°C for 44 h, and MTT assay was performed. (A), COS-7 cells. (B), HEp-2 cells. Data are presented as the mean ± SD of duplicate experiments. *p < 0.05.
Table 1.
Prediction of NLS sequences in functional proteins of S. aureus
Table 2.
Prediction of NLS sequences in hypothetical proteins of S. aureus
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