RT Journal Article
SR Electronic
T1 A cross–nearest neighbor/Monte Carlo algorithm for single-molecule localization microscopy defines interactions between p53, Mdm2, and MEG3
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 857912
DO 10.1101/857912
A1 Bauer, Nicholas C.
A1 Yang, Anli
A1 Wang, Xin
A1 Zhou, Yunli
A1 Klibanski, Anne
A1 Soberman, Roy J.
YR 2021
UL http://biorxiv.org/content/early/2021/04/11/857912.abstract
AB The functions of long noncoding (lnc)RNAs such as MEG3 are defined by their interactions with other RNAs and proteins. These interactions, in turn, are shaped by their subcellular localization and temporal context. Therefore, it is important to be able to analyze the relationships of lncRNAs while preserving cellular architecture. The ability of MEG3 to suppress cell proliferation led to its recognition as a tumor suppressor. MEG3 has been proposed to activate p53 by disrupting the interaction of p53 with Mdm2. To test this mechanism in the native cellular context, we employed two-color direct stochastic optical reconstruction microscopy (dSTORM), a single-molecule localization microscopy (SMLM) technique to detect and quantify the localizations of p53, Mdm2, and MEG3 in U2OS cells. We developed a new cross-nearest neighbor/Monte Carlo algorithm to quantify the association of these molecules. Proof of concept for our method was obtained by examining the association between FKBP1A and mTOR, MEG3 and p53, and Mdm2 and p53. In contrast to previous models, our data support a model in which MEG3 modulates p53 independently of the interaction with Mdm2.Competing Interest StatementThe authors have declared no competing interest.