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

8 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

Fingerprint

Nuclear Localization Signals
Nuclear Proteins
Helicobacter pylori
Organism Cloning
Bacterial Proteins
Computational Biology
Open Reading Frames
Proteins
Pathology
Urease
COS Cells
Protein Subunits
Mutant Proteins
Protein Sorting Signals
Green Fluorescent Proteins
Cell Nucleus
Cytoplasm
Bacteria

Keywords

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

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

Cite this

Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori. / Lee, Jung Hwa; Jun, So Hyun; Baik, Seung Chul; Kim, Deok Ryong; Park, Jae-Yong; Lee, Yong Seok; Choi, Chul Hee; Lee, Je Chul.

In: Journal of Microbiological Methods, Vol. 91, No. 3, 01.12.2012, p. 490-496.

Research output: Contribution to journalArticle

Lee, Jung Hwa ; Jun, So Hyun ; Baik, Seung Chul ; Kim, Deok Ryong ; Park, Jae-Yong ; Lee, Yong Seok ; Choi, Chul Hee ; Lee, Je Chul. / Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori. In: Journal of Microbiological Methods. 2012 ; Vol. 91, No. 3. pp. 490-496.
@article{11432f8e0b76443589f914f090ef332f,
title = "Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori",
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.",
keywords = "Bacterial pathogenesis, Bioinformatics, Gateway cloning system, Nuclear localization signal, Nuclear targeting protein",
author = "Lee, {Jung Hwa} and Jun, {So Hyun} and Baik, {Seung Chul} and Kim, {Deok Ryong} and Jae-Yong Park and Lee, {Yong Seok} and Choi, {Chul Hee} and Lee, {Je Chul}",
year = "2012",
month = "12",
day = "1",
doi = "10.1016/j.mimet.2012.10.004",
language = "English",
volume = "91",
pages = "490--496",
journal = "Journal of Microbiological Methods",
issn = "0167-7012",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori

AU - Lee, Jung Hwa

AU - Jun, So Hyun

AU - Baik, Seung Chul

AU - Kim, Deok Ryong

AU - Park, Jae-Yong

AU - Lee, Yong Seok

AU - Choi, Chul Hee

AU - Lee, Je Chul

PY - 2012/12/1

Y1 - 2012/12/1

N2 - 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.

AB - 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.

KW - Bacterial pathogenesis

KW - Bioinformatics

KW - Gateway cloning system

KW - Nuclear localization signal

KW - Nuclear targeting protein

UR - http://www.scopus.com/inward/record.url?scp=84868253850&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84868253850&partnerID=8YFLogxK

U2 - 10.1016/j.mimet.2012.10.004

DO - 10.1016/j.mimet.2012.10.004

M3 - Article

C2 - 23079023

AN - SCOPUS:84868253850

VL - 91

SP - 490

EP - 496

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

SN - 0167-7012

IS - 3

ER -