Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori

Jung Hwa Lee, So Hyun Jun, Seung Chul Baik, Deok Ryong Kim, Jae Yong Park, Yong Seok Lee, Chul Hee Choi, Je Chul Lee

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Host cell pathology induced by nuclear targeting of bacterial proteins has recently been identified as a pathogenic mechanism of bacteria. However, very few bacterial proteins were identified to target the nuclei of host cells. This study was designed to screen nuclear targeting proteins with nuclear localization signals (NLSs) in Helicobacter pylori using a combination of bioinformatic analysis and the Gateway recombinational cloning system. Forty-nine functional or hypothetical proteins were predicted to carry the putative NLSs among 1570 open reading frames (ORFs) of H. pylori 26695. Entire sets of 49 H. pylori ORFs were cloned for the generation of green fluorescent protein-tagged proteins using the Gateway recombinational cloning system. Twenty-six H. pylori proteins with the putative NLSs were found to target in the nuclei of COS-7 cells, whereas 23 were localized in the cytoplasm of host cells. Deletion of NLS sequences from four selected nuclear targeting proteins, urease subunit A, Omp18, secreted protein involved in flagellar motility, and response regulator, resulted in cytoplasmic localization of these mutant proteins. In conclusion, a combination of bioinformatic analysis and the Gateway cloning system was shown to be a useful tool for large-scale screening of nuclear targeting proteins with NLSs in H. pylori, which can be used to better understand the H. pylori-directed host cell pathology.

Original languageEnglish
Pages (from-to)490-496
Number of pages7
JournalJournal of Microbiological Methods
Volume91
Issue number3
DOIs
Publication statusPublished - 2012 Dec 1
Externally publishedYes

Keywords

  • Bacterial pathogenesis
  • Bioinformatics
  • Gateway cloning system
  • Nuclear localization signal
  • Nuclear targeting protein

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

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