 |
 Previous Article | Next Article 
Journal of Neuroscience, Vol 6, 1004-1012, Copyright © 1986 by Society for Neuroscience
Interval between the synthesis and assembly of cytoskeletal proteins in cultured neurons
MM Black, P Keyser and E Sobel
We have used pulse-chase experiments to study the time interval between the
synthesis and assembly of tubulin and neurofilament proteins (NFP) in
sympathetic neurons grown in tissue culture. After varying pulse- chase
times, cultures were extracted with Triton X-100 such that polymerized
tubulin and NFP were insoluble, while unassembled tubulin and NFP were
quantitatively solubilized. The partitioning of labeled tubulin and NFP
between Triton X-100-soluble and insoluble, or cytoskeletal, fractions was
determined with an isoelectric focusing X SDS gel electrophoresis assay.
Labeled tubulin and NFP in cultures pulse-labeled for 5-10 min partitions
primarily with the soluble fraction. When pulse-labeled cultures were
chased for increasing periods of time, relatively more of the total labeled
tubulin and NFP partitioned with the cytoskeleton, attaining maximal values
after chase times of 60-120 and 15-30 min, respectively. The maximal values
for the relative levels of labeled tubulin and NFP in polymer were 70-75
and greater than 90%, respectively. The levels of labeled tubulin and NFP
synthesized during a short pulse-label remained constant for at least 2 hr,
indicating that selective turnover of soluble tubulin and NFP does not
detectably contribute to the changes in solubility properties of these
proteins observed in the pulse-chase experiments. These results indicate
that newly synthesized tubulin and NFP are rapidly assembled from soluble
precursors. The lag between the synthesis and assembly of the
145,000-molecular-weight NFP is not related to its phosphorylation because
its initial incorporation into the cytoskeleton occurs prior to its
phosphorylation.(ABSTRACT TRUNCATED AT 250 WORDS)
This article has been cited by other articles:
|
|
|
|
 
T. Alkam, A. Nitta, H. Mizoguchi, A. Itoh, R. Murai, T. Nagai, K. Yamada, and T. Nabeshima
The Extensive Nitration of Neurofilament Light Chain in the Hippocampus Is Associated with the Cognitive Impairment Induced by Amyloid {beta} in Mice
J. Pharmacol. Exp. Ther.,
October 1, 2008;
327(1):
137 - 147.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Fasani, A. Bocquet, P. Robert, A. Peterson, and J. Eyer
The amount of neurofilaments aggregated in the cell body is controlled by their increased sensitivity to trypsin-like proteases
J. Cell Sci.,
February 22, 2004;
117(6):
861 - 869.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Bacher, B. Nickel, P. Emig, U. Vanhoefer, S. Seeber, A. Shandra, T. Klenner, and T. Beckers
D-24851, a Novel Synthetic Microtubule Inhibitor, Exerts Curative Antitumoral Activity in Vivo, Shows Efficacy toward Multidrug-resistant Tumor Cells, and Lacks Neurotoxicity
Cancer Res.,
January 1, 2001;
61(1):
392 - 399.
[Abstract]
[Full Text]
|
|
|
|
|
|
|
S. Roy, P. Coffee, G. Smith, R. K. H. Liem, S. T. Brady, and M. M. Black
Neurofilaments Are Transported Rapidly But Intermittently in Axons: Implications for Slow Axonal Transport
J. Neurosci.,
September 15, 2000;
20(18):
6849 - 6861.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. J. Ahmad, W. Yu, F. J. McNally, and P. W. Baas
An Essential Role for Katanin in Severing Microtubules in the Neuron
J. Cell Biol.,
April 19, 1999;
145(2):
305 - 315.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
T. J. Koehnle and A. Brown
Slow Axonal Transport of Neurofilament Protein in Cultured Neurons
J. Cell Biol.,
February 8, 1999;
144(3):
447 - 458.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
Q. Zhu, M. Lindenbaum, F. Levavasseur, H. Jacomy, and J.-P. Julien
Disruption of the NF-H Gene Increases Axonal Microtubule Content and Velocity of Neurofilament Transport: Relief of Axonopathy Resulting from the Toxin {beta},{beta}'-Iminodipropionitrile
J. Cell Biol.,
October 5, 1998;
143(1):
183 - 193.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. J. Ahmad, C. J. Echeverri, R. B. Vallee, and P. W. Baas
Cytoplasmic Dynein and Dynactin Are Required for the Transport of Microtubules into the Axon
J. Cell Biol.,
January 26, 1998;
140(2):
391 - 401.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A Brown
Contiguous phosphorylated and non-phosphorylated domains along axonal neurofilaments
J. Cell Sci.,
January 2, 1998;
111(4):
455 - 467.
[Abstract]
[PDF]
|
|
|
|
|
|
|
F. Ahmad and P. Baas
Microtubules released from the neuronal centrosome are transported into the axon
J. Cell Sci.,
January 8, 1995;
108(8):
2761 - 2769.
[Abstract]
[PDF]
|
|
|
|
|
|
|
A Roobol, F. Holmes, N. Hayes, A. Baines, and M. Carden
Cytoplasmic chaperonin complexes enter neurites developing in vitro and differ in subunit composition within single cells
J. Cell Sci.,
January 4, 1995;
108(4):
1477 - 1488.
[Abstract]
[PDF]
|
|
|
|
|
|
|
C. Chou, C. Riopel, L. Rott, and M. Omary
A significant soluble keratin fraction in 'simple' epithelial cells. Lack of an apparent phosphorylation and glycosylation role in keratin solubility
J. Cell Sci.,
January 6, 1993;
105(2):
433 - 444.
[Abstract]
[PDF]
|
|
|
|
|
