Visualizing Transcription Factor Binding on Mitotic Chromosomes Using Single-Molecule Live-Cell Imaging.
Methods Mol Biol
; 2038: 239-250, 2019.
Article
em En
| MEDLINE
| ID: mdl-31407289
ABSTRACT
For over two decades, scientists have observed that most transcription factors (TFs) become excluded from mitotic chromosomes of mammalian cells undergoing cell division. The few TFs that were observed to remain bound to chromosomes have been termed mitotic bookmarkers and were predicted to play important roles in reestablishing transcription after mitosis. Using live-cell imaging of endogenous TFs in mouse embryonic stem cells, we discovered that the observed exclusion from mitotic chromosomes is largely a result of formaldehyde cross-linking and that in fact, most TFs bind to mitotic chromosomes throughout mitosis. Here, we describe the single-molecule live-cell imaging and analytical tools we used to characterize and quantify TF diffusion and binding as mouse embryonic stem cells proceed through mitosis.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Fatores de Transcrição
/
Cromossomos de Mamíferos
/
Imagem Molecular
/
Células-Tronco Embrionárias Murinas
/
Imagem Individual de Molécula
/
Microscopia de Fluorescência
/
Mitose
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
Canadá