Screening and identification of natural antisense transcripts in Helicobacter pylori by a novel approach based on RNase I protection assay.
Natural antisense transcripts (NATs) are endogenous RNA molecules that exhibit partial or complete complementarity to other RNAs. Studies have shown that NATs may participate in a broad range of gene regulatory events. The identification of NATs in human, mouse and Escherichia coli has revealed their widespread occurrence in both eukaryotic and prokaryotic life. However, little is known about NATs in Helicobacter pylori (H. pylori), a human pathogen which is associated with gastric diseases. Here we systematically screened NATs in H. pylori by a novel experimental strategy based on RNase I protection assay. We successfully constructed a cDNA library of NATs and developed a novel poly(A)-tailed RT-PCR method to monitor the expression of NATs. After sequencing, bioinformatic analysis and expression detection, two novel NATs (NAT-39 and NAT-67) were confirmed. They were, respectively, complementary to the following genes: iron-regulated outer membrane protein (frpB) and periplasmic iron-binding protein (ceuE). Taken together, the results suggest that NAT-39 and NAT-67 may participate in the regulation of iron homeostasis in H. Pylori in a sequence complementary manner with target mRNAs.
Xiao
Bin
B
Department of Clinical Microbiology and Immunology, Faculty of Medical Laboratory Science, Third Military Medical University, Chongqing, People's Republic of China.
Li
Wei
W
Guo
Gang
G
Li
Bo-Sheng
BS
Liu
Zhen
Z
Tang
Bin
B
Mao
Xu-Hu
XH
Zou
Quan-Ming
QM
D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D016680:Genome, Bacterial / Q000235:genetics; D016480:Helicobacter pylori / Q000235:genetics*; D008969:Molecular Sequence Data; D023282:Nuclease Protection Assays / Q000379:methods*; D009690:Nucleic Acid Conformation; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*
D016372:RNA, Antisense; D012333:RNA, Messenger; D012259:Ribonuclease, Pancreatic