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Monitoring of chromatin organization in live cells by FRIC. Effects of the inner nuclear membrane protein Samp1.

Abstract：

:In most cells, transcriptionally inactive heterochromatin is preferentially localized in the nuclear periphery and transcriptionally active euchromatin is localized in the nuclear interior. Different cell types display characteristic chromatin distribution patterns, which change dramatically during cell differentiation, proliferation, senescence and different pathological conditions. Chromatin organization has been extensively studied on a cell population level, but there is a need to understand dynamic reorganization of chromatin at the single cell level, especially in live cells. We have developed a novel image analysis tool that we term Fluorescence Ratiometric Imaging of Chromatin (FRIC) to quantitatively monitor dynamic spatiotemporal distribution of euchromatin and total chromatin in live cells. A vector (pTandemH) assures stoichiometrically constant expression of the histone variants Histone 3.3 and Histone 2B, fused to EGFP and mCherry, respectively. Quantitative ratiometric (H3.3/H2B) imaging displayed a concentrated distribution of heterochromatin in the periphery of U2OS cell nuclei. As proof of concept, peripheral heterochromatin responded to experimental manipulation of histone acetylation. We also found that peripheral heterochromatin depended on the levels of the inner nuclear membrane protein Samp1, suggesting an important role in promoting peripheral heterochromatin. Taken together, FRIC is a powerful and robust new tool to study dynamic chromatin redistribution in live cells.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Bergqvist C,Niss F,Figueroa RA,Beckman M,Maksel D,Jafferali MH,Kulyté A,Ström AL,Hallberg E

doi

10.1093/nar/gkz123

subject

Has Abstract

pub_date

2019-05-21 00:00:00

pages

e49

issue

9

eissn

0305-1048

issn

1362-4962

pii

5356937

journal_volume

47

pub_type

杂志文章

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