title: Cloning of a novel protein interacting with BRS-3 and its effects in wound repair of bronchial epithelial cells.
abstract: Bombesin receptor subtype 3 (BRS-3), the orphan bombesin receptor, may play a role in the regulation of stress responses in lung and airway epithelia. Bombesin receptor activated protein (BRAP )is a novel protein we found in our previous study which interacts with BRS-3. This study was designed to observe the subcellular location and wound repair function of BRAP in human bronchial epithelial cells (HBECs). BRAP ORF was amplified by RT-PCR and ligated to pEGFP-C1 vector, and then the recombinant plasmid pEGFP-C1-BRAP was transfected into Hela cells. The location of BRAP protein was observed by laser confocal microscope, and the expression of it was analyzed by Western-blot. At the same time,we built the recombinant plasmid pcDNA3.1(+)-BRAP, transfected it into HBECs and observed its impact on cell cycle and wound repair of HBECs. The results showed that BRAP locates in membrane and cytoplasm and increases significantly in transfected cells. Flow cytometry results demonstrated that the recombinant plasmid increases S phase plus G2 phase of cell cycle by 25%. Microscopic video analysis system showed that the repair index of wounded HBECs increases by 20% through stable expression of BRAP. The present study demonstrated that BRAP locates in the membrane and cytoplasm, suggesting that this protein is a cytoplasm protein, which promotes cell cycle and wound repair of HBECs.
authors.affiliation: Department of Physiology, Xiangya School of Medicine, Central South University, Changsha, China.
authors.forename: Hui Jun
authors.initials: HJ
authors.lastname: Liu
authors.forename: Yu Rong
authors.initials: YR
authors.lastname: Tan
authors.forename: Meng Lan
authors.initials: ML
authors.lastname: Li
authors.forename: Chi
authors.initials: C
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Xiang
authors.forename: Xiao Qun
authors.initials: XQ
authors.lastname: Qin
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D005434:Flow Cytometry; D016195:G2 Phase; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D018004:Receptors, Bombesin / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase; D014162:Transfection; D014945:Wound Healing
chemical_list: C519991:C6orf89 protein, human; D011506:Proteins; D018004:Receptors, Bombesin; D011993:Recombinant Fusion Proteins; C107120:bombesin receptor subtype 3; D049452:Green Fluorescent Proteins