3D Protein Dynamics in the Cell Nucleus

详细信息    查看全文

• 作者：Anand P. Singh¹ ; Ré ; mi Galland² ; ³ ; Megan L. Finch-Edmondson¹ ; ⁴ ; Gianluca Grenci¹ ; Jean-Baptiste Sibarita² ; ³ ; Vincent Studer² ; ³ ; Virgile Viasnoff¹ ; ⁵ ; ⁶ ; ^{virgile.viasnoff@espci.fr} ; Timothy E. Saunders¹ ; ⁵ ; ⁷ ; ^{dbsste@nus.edu.sg}
• 刊名：Biophysical Journal
• 出版年：2017
• 出版时间：10 January 2017
• 年：2017
• 卷：112
• 期：1
• 页码：133-142
• 全文大小：2616 K
• 卷排序：112

文摘

The three-dimensional (3D) architecture of the cell nucleus plays an important role in protein dynamics and in regulating gene expression. However, protein dynamics within the 3D nucleus are poorly understood. Here, we present, to our knowledge, a novel combination of 1) single-objective based light-sheet microscopy, 2) photoconvertible proteins, and 3) fluorescence correlation microscopy, to quantitatively measure 3D protein dynamics in the nucleus. We are able to acquire >3400 autocorrelation functions at multiple spatial positions within a nucleus, without significant photobleaching, allowing us to make reliable estimates of diffusion dynamics. Using this tool, we demonstrate spatial heterogeneity in Polymerase II dynamics in live U2OS cells. Further, we provide detailed measurements of human-Yes-associated protein diffusion dynamics in a human gastric cancer epithelial cell line.