RT Journal Article
SR Electronic
T1 Time-resolved structured illumination microscopy reveals key principles of Xist RNA spreading
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.11.24.396473
DO 10.1101/2020.11.24.396473
A1 Lisa Rodermund
A1 Heather Coker
A1 Roel Oldenkamp
A1 Guifeng Wei
A1 Joseph Bowness
A1 Bramman Rajkumar
A1 Tatyana Nesterova
A1 David Pinto
A1 Lothar Schermelleh
A1 Neil Brockdorff
YR 2021
UL http://biorxiv.org/content/early/2021/04/26/2020.11.24.396473.abstract
AB Xist RNA directs the process of X-chromosome inactivation in mammals by spreading in cis along the chromosome from which it is transcribed and recruiting chromatin modifiers to silence gene transcription. To elucidate mechanisms of Xist RNA cis-confinement, we established a sequential dual color labeling, super-resolution imaging approach to trace individual Xist RNA molecules over time, enabling us to define fundamental parameters of spreading. We demonstrate a feedback mechanism linking Xist RNA synthesis and degradation, and an unexpected physical coupling between preceding and newly synthesized Xist RNA molecules. Additionally, we show that the protein SPEN, a key factor for Xist-mediated gene-silencing, has a distinct function in Xist RNA localization, stability and in coupling behavior. Our results provide important insights towards understanding the unique dynamic properties of Xist RNA.One Sentence Summary Visualizing Xist RNA dynamics in single cells during X chromosome inactivationCompeting Interest StatementThe authors have declared no competing interest.