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Media type:
E-Article
Title:
Biophysical characterisation of human LincRNA-p21 sense and antisense Alu inverted repeats
Contributor:
D’Souza, Michael H;
Mrozowich, Tyler;
Badmalia, Maulik D;
Geeraert, Mitchell;
Frederickson, Angela;
Henrickson, Amy;
Demeler, Borries;
Wolfinger, Michael T;
Patel, Trushar R
Published:
Oxford University Press (OUP), 2022
Published in:
Nucleic Acids Research, 50 (2022) 10, Seite 5881-5898
Language:
English
DOI:
10.1093/nar/gkac414
ISSN:
0305-1048;
1362-4962
Origination:
Footnote:
Description:
AbstractHuman 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.