Prediction and screening of nuclear targeting proteins with nuclear localization signals in Helicobacter pylori.
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.
Department of Microbiology, Kyungpook National University School of Medicine, Jung-gu, Daegu, Republic of Korea.
Jung Hwa
JH
Lee
So Hyun
SH
Jun
Seung Chul
SC
Baik
Deok Ryong
DR
Kim
Jae-Yong
JY
Park
Yong Seok
YS
Lee
Chul Hee
CH
Choi
Je Chul
JC
Lee
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D019295:Computational Biology / Q000379:methods*; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism; D016480:Helicobacter pylori / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019913:Nuclear Localization Signals; D021381:Protein Transport; D011995:Recombination, Genetic
D001426:Bacterial Proteins; D019913:Nuclear Localization Signals