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Helicobacter pylori HP 0425 Targets the Nucleus with DN ase I‐Like Activity
Author(s) -
Kim JungMin,
Choe MinHo,
Asaithambi Killivalavan,
Song JaeYoung,
Lee Yong Seok,
Lee Je Chul,
Seo JiHyun,
Kang HyungLyun,
Lee Kon Ho,
Lee WooKon,
Cho MyungJe,
Rhee KwangHo,
Youn HeeShang,
Baik SeungChul
Publication year - 2016
Publication title -
helicobacter
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.206
H-Index - 79
eISSN - 1523-5378
pISSN - 1083-4389
DOI - 10.1111/hel.12271
Subject(s) - microbiology and biotechnology , chemistry , biology
Background and Aims Nuclear targeting of bacterial proteins has a significant impact on host cell pathology. Helicobacter pylori have many nuclear targeting proteins that translocate into the nucleus of host cells. H. pylori HP 0425, annotated as hypothetical, has a nuclear localization signal ( NLS ) sequence, but its function has not been demonstrated. The aim of this experiment was to address the nuclear translocation of HP 0425 and determine the effect of HP 0425 pathology on host cells. Materials and Methods To investigate the nuclear localization of HP 0425, it was expressed in AGS and MKN ‐1 cells as a GFP fusion protein ( pEGFP – HP 0425), and its localization was analyzed by confocal microscopy. Recombinant HP 0425 ( rHP 0425) protein was overproduced as a GST fusion protein in Escherichia coli and purified by glutathione‐affinity column chromatography. Purified rHP 0425 was examined for cytotoxicity and DN ase activity. Results The pEGFP – HP 0425 fluorescence was expressed in the nucleus and cytosol fraction of cells, while it was localized in the cytoplasm in the negative control. This protein exhibited DN ase activity under various conditions, with the highest DN ase activity in the presence of manganese. In addition, the rHP 0425 protein efficiently decreased cell viability in a concentration‐dependent manner. Conclusions These results suggest that HP 0425 carrying a nuclear localization signal sequence translocates into the nucleus of host cells and degrades genomic DNA by DN ase I‐like enzymatic activity, which is a new pathogenic strategy of H. pylori in the host.