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

Bergqvist, Cecilia; Niss, Frida; Figueroa, Ricardo A; Beckman, Marie; Maksel, Danuta; Jafferali, Mohammed H; Kulyté, Agné; Ström, Anna-Lena; Hallberg, Einar

Bergqvist, Cecilia; Niss, Frida; Figueroa, Ricardo A; Beckman, Marie; Maksel, Danuta; Jafferali, Mohammed H; Kulyté, Agné; Ström, Anna-Lena; Hallberg, Einar.

Afiliación

Bergqvist C; Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius väg 16B, SE-106 91 Stockholm, Sweden.
Niss F; Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius väg 16B, SE-106 91 Stockholm, Sweden.
Figueroa RA; Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius väg 16B, SE-106 91 Stockholm, Sweden.
Beckman M; Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius väg 16B, SE-106 91 Stockholm, Sweden.
Maksel D; Institute of Environmental Medicine, Karolinska Institutet SE-171 77 Sweden.
Jafferali MH; Monash Molecular Crystallisation Facility (MMCF), Monash University, VIC 3800, Australia.
Kulyté A; Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius väg 16B, SE-106 91 Stockholm, Sweden.
Ström AL; Lipid laboratory, Department of Medicine, Karolinska Institutet, SE-141 57 Huddinge, Sweden.
Hallberg E; Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius väg 16B, SE-106 91 Stockholm, Sweden.

Nucleic Acids Res ; 47(9): e49, 2019 05 21.

Article en En | MEDLINE | ID: mdl-30793190

RESUMEN

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.

Asunto(s)

Cromatina/genética; Proteínas de la Membrana/genética; Imagen Molecular/métodos; Proteínas Nucleares/genética; Acetilación; Línea Celular; Núcleo Celular/genética; Eucromatina/genética; Heterocromatina/genética; Histonas/genética; Humanos; Membrana Nuclear/genética; Procesamiento Proteico-Postraduccional/genética

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Nucleares / Cromatina / Imagen Molecular / Proteínas de la Membrana Límite: Humans Idioma: En Revista: Nucleic Acids Res Año: 2019 Tipo del documento: Article País de afiliación: Suecia Pais de publicación: Reino Unido

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