LRP16 gene function based on bioinformatic analysis
10.3321/j.issn:1000-467X.2009.12.010
- VernacularTitle:基于生物信息学分析人类LRP16基因功能初步研究
- Author:
Yang BO
1
;
Lu XUE-CHUN
;
Chi XIAO-HUA
;
Han WEI-DONG
;
Yu LI
;
Lou FANG-DING
Author Information
1. 解放军总医院
- Keywords:
LRP16;
function;
bioinformatics
- From:Chinese Journal of Cancer
2009;28(12):1283-1290
- CountryChina
- Language:Chinese
-
Abstract:
Background and Objective:LRP16 is a human novel gene linked to leukemia identified recently.However. its biological function is not fully clarified so far.This study was to investigate the biological function of human LRP16 gene by database-aided bioinformatics analysis. Methods:The structures and functions of LRP16 gene promoter and its coding protein were analyzed using bioinformatics prediction,and further experimental testing was performed.The recombinants of pGL3-basic and LRP16 promoter subclones were constructed for luciferase activity analysis.The recombinant of LRP16 open reading frame coding sequence and pcDNA3.1 eukaryotic expression vector was established and transfected into HL-60 and K562 cell lines.DNA damage of HL-60 cells after ultraviolet irradiation was evaluated using single cell gel electrophoresis. Cell cycle of K562 cells was analyzed by flow cytometry. Results:LRP16 promoter was a typical class Ⅱ eukaryotic promoter and its core regulation sequence was located within upstream -600 bp of transcriptional start site.In addition,seven cis-acting elements,which may be implicated in cell cycle,hematopoiesis regulation, cell proliferation and repair of DNA damage,were identified.Long type LRP16 coding protein contained homologous sequences of hismacro, COG2110,and A1PP with human histone H2A1C between 148 and 315 amino acid residue.The number of comet cells and the length of comet tail in HL-60 cells irradiated were significantly decreased and the number of living cell was significantly increased in LRP16-overexpression group compared with empty plasmid control group.The proliferation rate and ratio or quantity of G_2/M and S phases were significantly increased in LRP16-overexpression K562 group compared with empty plasmid control group.LRP16-overexpression in K562 cells promoted the transition of G_1 to S phase and plateau phase of cell proliferation was advanced.Conclusions:Promoter regulation prediction and protein domain analysis based on bioinformatics contribute to the study of gene function.LRP16 may play an important role in leukemia progression by promoting cell proliferation,regulating cell cycle,and antagonizing radiationinduced DNA damage.
