Balbiani ring mRNPs diffuse through and bind to clusters of large intranuclear molecular structures.
A detailed conception of intranuclear messenger ribonucleoprotein particle (mRNP) dynamics is required for the understanding of mRNP processing and gene expression outcome. We used complementary state-of-the-art fluorescence techniques to quantify native mRNP mobility at the single particle level in living salivary gland cell nuclei. Molecular beacons and fluorescent oligonucleotides were used to specifically label BR2.1 mRNPs by an in vivo fluorescence in situ hybridization approach. We characterized two major mobility components of the BR2.1 mRNPs. These components with diffusion coefficients of 0.3 ± 0.02 μm²/s and 0.73 ± 0.03 μm²/s were observed independently of the staining method and measurement technique used. The mobility analysis of inert tracer molecules revealed that the gland cell nuclei contain large molecular nonchromatin structures, which hinder the mobility of large molecules and particles. The mRNPs are not only hindered by these mobility barriers, but in addition also interact presumably with these structures, what further reduces their mobility and effectively leads to the occurrence of the two diffusion coefficients. In addition, we provide evidence that the remarkably high mobility of the large, 50 nm-sized BR2.1 mRNPs was due to the absence of retarding chromatin.
Institute for Physical and Theoretical Chemistry, Rheinische Friedrich-Wilhelms-University Bonn, Germany. veith@pc.uni-bonn.de
Roman
R
Veith
Thomas
T
Sorkalla
Eugen
E
Baumgart
Johannes
J
Anzt
Hanns
H
Häberlein
Sanjay
S
Tyagi
Jan Peter
JP
Siebrasse
Ulrich
U
Kubitscheck
D001483:Base Sequence; D059045:Chromosomal Puffs / Q000737:chemistry / Q000378:metabolism*; D004058:Diffusion; D006367:HeLa Cells; D006801:Humans; D008853:Microscopy; D009068:Movement; D011485:Protein Binding; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015347:RNA Probes / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013997:Time Factors
D015347:RNA Probes; D012261:Ribonucleoproteins; C026452:messenger ribonucleoprotein; D012313:RNA