期刊
NUCLEIC ACIDS RESEARCH
卷 50, 期 10, 页码 5881-5898出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkac414
关键词
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资金
- NSERC
- South Alberta Light Horse Regimental Foundation
- King's Own Calgary Regiment (RCAC) Funds Foundation
- Canadian Armed Forces Individual Learning Plan
- Alberta Innovates grant
- Natural Sciences and Engineering Research Council (NSERC) PGS-D award
- NSERC CGS-D award
- Canada Foundation for Innovation [CFI-37589, CFI 37155, CFI-41008]
- NSERC RTI [RTI 2020-00090]
- Canada 150 Research Chairs program [C150-2017-00015]
- National Institutes of Health [1R01GM120600]
- Canadian Natural Science and Engineering Research Council [DG-RGPIN-201905637]
- NSF/XSEDE [TG-MCB070039N]
- DIAMOND Synchrotron, UK [SM26855]
This study investigated the structure of human LincRNA-p21 using various techniques, determining its low-resolution three-dimensional structure and calculating high-resolution models. The study presents a new approach for understanding the structure of long noncoding RNAs.
Human Long Intergenic Noncoding RNA-p21 (LincRNA-p21) is a regulatory noncoding RNA that plays an important role in promoting apoptosis. LincRNA-p21 is also critical in down-regulating many p53 target genes through its interaction with a p53 repressive complex. The interaction between LincRNA-p21 and the repressive complex is likely dependent on the RNA tertiary structure. Previous studies have determined the two-dimensional secondary structures of the sense and antisense human LincRNA-p21 AluSx1 IRs using SHAPE. However, there were no insights into its three-dimensional structure. Therefore, we in vitro transcribed the sense and antisense regions of LincRNA-p21 AluSx1 Inverted Repeats (IRs) and performed analytical ultracentrifugation, size exclusion chromatography, light scattering, and small angle X-ray scattering (SAXS) studies. Based on these studies, we determined low-resolution, three-dimensional structures of sense and antisense LincRNA-p21. By adapting previously known two-dimensional information, we calculated their sense and antisense high-resolution models and determined that they agree with the low-resolution structures determined using SAXS. Thus, our integrated approach provides insights into the structure of LincRNA-p21 Alu IRs. Our study also offers a viable pipeline for combining the secondary structure information with biophysical and computational studies to obtain high-resolution atomistic models for long noncoding RNAs.
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