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 language | English |
|---|---|
| Pages (from-to) | 490-496 |
| Number of pages | 7 |
| Journal | Journal of Microbiological Methods |
| Volume | 91 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2012 Dec 1 |
| Externally published | Yes |
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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 journal › Article
}
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 -