Biogenesis of short intronic repeat 27nt small RNA from endothelial nitric oxide synthase gene Export

@article{citeulike:2630989, abstract = {Endothelial nitric oxide synthase (eNOS) is a constitutively expressed gene in endothelium that produces NO and critical for vascular integrity. Previously, we reported that the 27nt repeat polymorphism in eNOS intron 4 - a source of 27nt small RNA, which inhibits eNOS expression, were associated with cardiovascular risk and expression of the eNOS gene. In the current study, we investigated the biogenesis of the intron 4-derived 27nt small RNA. Using Northern blot, we showed that the eNOS-derived 27nt short intronic repeat RNA (sir-RNA) expressed only in the eNOS expressing endothelial cells. Cells containing 10x27nt or 5x27nt repeats produced higher levels of 27nt sir-RNA and lower levels of eNOS mRNA than the cells with 4x27nt repeats. The 27nt sir-RNA was mostly present within the endothelial nuclei. When the splicing junctions of the 27nt repeat containing intron 4 in the full-length eNOS cDNA vector were mutated, 27nt sir-RNA biogenesis was abolished. Suppression of Drosha or Dicer diminished the biogenesis of the 27nt sir-RNA. Our study suggests that the 27nt sir-RNA derived through eNOS pre-mRNA splicing may represent a new class of small RNA. The more eNOS is transcribed or higher number of the 27nt repeats, the more 27nt sir-RNA is produced, which functions as a negative feedback self-regulator by specifically inhibiting the host gene eNOS expression. This novel molecular model may be responsible for quantitative differences between individuals carrying different numbers of the polymorphic repeats hence the cardiovascular risk. 10.1074/jbc.M801933200}, author = {Zhang, Ming-Xiang and Zhang, Cheng and Shen, Ying H. and Wang, Jian and Li, Xiao N. and Zhang, Yun and Coselli, Joseph and Wang, Xing L.}, citeulike-article-id = {2630989}, citeulike-linkout-0 = {http://dx.doi.org/10.1074/jbc.M801933200}, day = {3}, doi = {10.1074/jbc.M801933200}, journal = {J. Biol. Chem.}, keywords = {no, rnai}, month = {April}, pages = {M801933200+}, posted-at = {2008-04-05 03:36:09}, priority = {2}, title = {Biogenesis of short intronic repeat 27nt small RNA from endothelial nitric oxide synthase gene}, url = {http://dx.doi.org/10.1074/jbc.M801933200}, year = {2008} }

TY - JOUR ID - citeulike:2630989 L3 - citeulike-article-id:2630989 N2 - Endothelial nitric oxide synthase (eNOS) is a constitutively expressed gene in endothelium that produces NO and critical for vascular integrity. Previously, we reported that the 27nt repeat polymorphism in eNOS intron 4 - a source of 27nt small RNA, which inhibits eNOS expression, were associated with cardiovascular risk and expression of the eNOS gene. In the current study, we investigated the biogenesis of the intron 4-derived 27nt small RNA. Using Northern blot, we showed that the eNOS-derived 27nt short intronic repeat RNA (sir-RNA) expressed only in the eNOS expressing endothelial cells. Cells containing 10x27nt or 5x27nt repeats produced higher levels of 27nt sir-RNA and lower levels of eNOS mRNA than the cells with 4x27nt repeats. The 27nt sir-RNA was mostly present within the endothelial nuclei. When the splicing junctions of the 27nt repeat containing intron 4 in the full-length eNOS cDNA vector were mutated, 27nt sir-RNA biogenesis was abolished. Suppression of Drosha or Dicer diminished the biogenesis of the 27nt sir-RNA. Our study suggests that the 27nt sir-RNA derived through eNOS pre-mRNA splicing may represent a new class of small RNA. The more eNOS is transcribed or higher number of the 27nt repeats, the more 27nt sir-RNA is produced, which functions as a negative feedback self-regulator by specifically inhibiting the host gene eNOS expression. This novel molecular model may be responsible for quantitative differences between individuals carrying different numbers of the polymorphic repeats hence the cardiovascular risk. 10.1074/jbc.M801933200 JF - J. Biol. Chem. TI - Biogenesis of short intronic repeat 27nt small RNA from endothelial nitric oxide synthase gene SP - M801933200 KW - no KW - rnai AU - Zhang, Ming-Xiang AU - Zhang, Cheng AU - Shen, Ying AU - Wang, Jian AU - Li, Xiao AU - Zhang, Yun AU - Coselli, Joseph AU - Wang, Xing PY - 2008/04/03/ UR - http://dx.doi.org/10.1074/jbc.M801933200 ER -