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  4. Spatial separation of Xist RNA and polycomb proteins revealed by superresolution microscopy
 
research article

Spatial separation of Xist RNA and polycomb proteins revealed by superresolution microscopy

Cerase, Andrea
•
Smeets, Daniel
•
Tang, Y. Amy
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2014
Proceedings Of The National Academy Of Sciences Of The United States Of America (PNAS)

In female mammals, one of the two X chromosomes is transcriptionally silenced to equalize X-linked gene dosage relative to XY males, a process termed X chromosome inactivation. Mechanistically, this is thought to occur via directed recruitment of chromatin modifying factors by the master regulator, X-inactive specific transcript (Xist) RNA, which localizes in cis along the entire length of the chromosome. A well-studied example is the recruitment of polycomb repressive complex 2 (PRC2), for which there is evidence of a direct interaction involving the PRC2 proteins Enhancer of zeste 2 (Ezh2) and Supressor of zeste 12 (Suz12) and the A-repeat region located at the 5′ end of Xist RNA. In this study, we have analyzed Xist-mediated recruitment of PRC2 using two approaches, microarray-based epigenomic mapping and superresolution 3D structured illumination microscopy. Making use of an ES cell line carrying an inducible Xist transgene located on mouse chromosome 17, we show that 24 h after synchronous induction of Xist expression, acquired PRC2 binding sites map predominantly to gene-rich regions, notably within gene bodies. Paradoxically, these new sites of PRC2 deposition do not correlate with Xist-mediated gene silencing. The 3D structured illumination microscopy was performed to assess the relative localization of PRC2 proteins and Xist RNA. Unexpectedly, we observed significant spatial separation and absence of colocalization both in the inducible Xist transgene ES cell line and in normal XX somatic cells. Our observations argue against direct interaction between Xist RNA and PRC2 proteins and, as such, prompt a reappraisal of the mechanism for PRC2 recruitment in X chromosome inactivation.

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Type
research article
DOI
10.1073/pnas.1312951111
Author(s)
Cerase, Andrea
Smeets, Daniel
Tang, Y. Amy
Gdula, Michal
Kraus, Felix
Spivakov, Mikhail
Moindrot, Benoit
Leleu, Marion
Tattermusch, Anna
Demmerle, Justin
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Date Issued

2014

Published in
Proceedings Of The National Academy Of Sciences Of The United States Of America (PNAS)
Volume

111

Issue

6

Start page

2235

End page

2240

Editorial or Peer reviewed

REVIEWED

Written at

OTHER

EPFL units
SV  
Available on Infoscience
March 31, 2017
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/136174
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