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Mol Cell ; 67(2): 282-293.e7, 2017 Jul 20.
Artículo en Inglés | MEDLINE | ID: mdl-28712725

RESUMEN

The eukaryotic genome is organized within cells as chromatin. For proper information output, higher-order chromatin structures can be regulated dynamically. How such structures form and behave in various cellular processes remains unclear. Here, by combining super-resolution imaging (photoactivated localization microscopy [PALM]) and single-nucleosome tracking, we developed a nuclear imaging system to visualize the higher-order structures along with their dynamics in live mammalian cells. We demonstrated that nucleosomes form compact domains with a peak diameter of ∼160 nm and move coherently in live cells. The heterochromatin-rich regions showed more domains and less movement. With cell differentiation, the domains became more apparent, with reduced dynamics. Furthermore, various perturbation experiments indicated that they are organized by a combination of factors, including cohesin and nucleosome-nucleosome interactions. Notably, we observed the domains during mitosis, suggesting that they act as building blocks of chromosomes and may serve as information units throughout the cell cycle.


Asunto(s)
Ensamble y Desensamble de Cromatina , Heterocromatina/metabolismo , Microscopía por Video/métodos , Mitosis , Nucleosomas/metabolismo , Animales , Proteínas de Ciclo Celular/metabolismo , Diferenciación Celular , Proteínas Cromosómicas no Histona/metabolismo , Cromosomas Humanos , Células HCT116 , Células HeLa , Heterocromatina/química , Humanos , Ratones , Movimiento (Física) , Conformación de Ácido Nucleico , Nucleosomas/química , Conformación Proteica , Interferencia de ARN , Relación Estructura-Actividad , Factores de Tiempo , Transcripción Genética , Transfección , Cohesinas
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