Abstract
Understanding gene expression requires the ability to follow the fate of individual molecules. Here we use a cellular system for monitoring messenger RNA (mRNA)expression to characterize the movement in real time of single mRNA-protein complexes (mRNPs) in the nucleus of living mammalian cells. This mobility was not directed but was governed by simple diffusion. Some mRNPs were partially corralled throughout the nonhomogenous nuclear environment, but no accumulation at subnuclear domains was observed. Following energy deprivation, energy-independent motion of mRNPs was observed in a highly ATP-dependent nuclear environment; movements were constrained to chromatin-poor domains and excluded by newly formed chromatin barriers. This observation resolves a controversy, showing that the energetic requirements of nuclear mRNP trafficking are consistent with a diffusional model.
MeSH Terms
Active Transport, Cell Nucleus
Adenosine Triphosphate/metabolism
Capsid Proteins/genetics,metabolism
Cell Line, Tumor
Cell Nucleus/metabolism
Chromatin/metabolism
Cytoplasm/metabolism
Diffusion
Energy Metabolism
Fluorescence Recovery After Photobleaching
Globins/genetics,metabolism
Green Fluorescent Proteins
Humans
In Situ Hybridization, Fluorescence
Luminescent Proteins/genetics,metabolism
Peroxisomes/metabolism
Protein Biosynthesis
Proteins/genetics,metabolism
RNA, Messenger/genetics,metabolism
RNA-Binding Proteins/genetics,metabolism
Recombinant Fusion Proteins/metabolism
Transcription, Genetic
Transfection
Chemicals
Capsid Proteins
Chromatin
Luminescent Proteins
Proteins
RNA, Messenger
RNA-Binding Proteins
Recombinant Fusion Proteins
Green Fluorescent Proteins
Adenosine Triphosphate
Globins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Shav-Tal Yaron
Departments of Anatomy and Structural Biology and Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Darzacq Xavier
Shenoy Shailesh M
Fusco Dahlene
Janicki Susan M
Spector David L
Singer Robert H
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