|
|
|
|
|
||
|
|||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | |||||
Journal of Cell Science, Vol 109, Issue 9 2299-2310, Copyright © 1996 by Company of Biologists
JOURNAL ARTICLES |
A Watakabe, R Kobayashi and DM Helfman
Cold Spring Harbor Laboratory, NY 11724, USA.
We have identified and characterized two proteins in rat brain that bind to the neuron-specific tropomyosin isoform, TMBr3. The two proteins were identified by blot overlay assay, in which the proteins immobilized on the membrane were probed by epitope-tagged TMBr3, followed by detection with anti-epitope antibody. We have purified these proteins using a TMBr3 affinity column. Peptide sequencing as well as immunoblotting showed that one of the two proteins is identical to tropomodulin, a tropomyosin-binding protein originally identified in erythrocytes. The cDNA for the other protein was cloned from an adult rat brain cDNA library using degenerate oligonucleotides that we designed based on the peptide sequences. Sequence analysis of the cDNA clone revealed this protein to be a novel isoform of tropomodulin which is the product of a distinct gene, and is herein referred to as N-tropomodulin. Recombinant N-tropomodulin bound to TMBr3 as well as to other low molecular mass tropomyosins (TM5a or TM5), but not to high molecular mass tropomyosins (TM2 or TMBr1). Northern blotting and RNase protection assays as well as immunoblotting showed that N-tropomodulin is expressed predominantly in brain. Furthermore, RNase protection assays revealed no alternatively spliced regions within the coding sequence. Developmentally, N-tropomodulin was detected in rat brain as early as embryonic day 14 and reaches the adult level before birth. Immunofluorescence of primary frontal cortex cell cultures showed that N-tropomodulin is specifically expressed in neurons. The neuron-specific expression of N-tropomodulin strongly suggests specialized roles of this TM-binding protein in neurons.
This article has been cited by other articles:
|
|
 |
|
  P. Gunning, G. O'neill, and E. Hardeman Tropomyosin-Based Regulation of the Actin Cytoskeleton in Time and Space Physiol Rev, January 1, 2008; 88(1): 1 - 35. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Y. Kong and L. Kedes Leucine 135 of Tropomodulin-1 Regulates Its Association with Tropomyosin, Its Cellular Localization, and the Integrity of Sarcomeres J. Biol. Chem., April 7, 2006; 281(14): 9589 - 9599. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. J. Greenfield, A. S. Kostyukova, and S. E. Hitchcock-DeGregori Structure and Tropomyosin Binding Properties of the N-Terminal Capping Domain of Tropomodulin 1 Biophys. J., January 1, 2005; 88(1): 372 - 383. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Y. Kong and L. Kedes Cytoplasmic Nuclear Transfer of the Actin-capping Protein Tropomodulin J. Biol. Chem., July 16, 2004; 279(29): 30856 - 30864. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. L. Fritz-Six, P. R. Cox, R. S. Fischer, B. Xu, C. C. Gregorio, H. Y. Zoghbi, and V. M. Fowler Aberrant myofibril assembly in tropomodulin1 null mice leads to aborted heart development and embryonic lethality J. Cell Biol., December 8, 2003; 163(5): 1033 - 1044. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. M. Fowler, N. J. Greenfield, and J. Moyer Tropomodulin Contains Two Actin Filament Pointed End-capping Domains J. Biol. Chem., October 10, 2003; 278(41): 40000 - 40009. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Chu, J. Chen, M. C. Reedy, C. Vera, K.-L. P. Sung, and L. A. Sung E-Tmod capping of actin filaments at the slow-growing end is required to establish mouse embryonic circulation Am J Physiol Heart Circ Physiol, May 1, 2003; 284(5): H1827 - H1838. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. G. Dos Remedios, D. Chhabra, M. Kekic, I. V. Dedova, M. Tsubakihara, D. A. Berry, and N. J. Nosworthy Actin Binding Proteins: Regulation of Cytoskeletal Microfilaments Physiol Rev, April 1, 2003; 83(2): 433 - 473. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. A. Conley Leiomodin and tropomodulin in smooth muscle Am J Physiol Cell Physiol, June 1, 2001; 280(6): C1645 - C1656. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. S. Fischer, A. Lee, and V. M. Fowler Tropomodulin and Tropomyosin Mediate Lens Cell Actin Cytoskeleton Reorganization In Vitro Invest. Ophthalmol. Vis. Sci., January 1, 2000; 41(1): 166 - 174. [Abstract] [Full Text]
|
|
|
|
|
|
A. Almenar-Queralt, A. Lee, C. A. Conley, L. R. de Pouplana, and V. M. Fowler Identification of a Novel Tropomodulin Isoform, Skeletal Tropomodulin, That Caps Actin Filament Pointed Ends in Fast Skeletal Muscle J. Biol. Chem., October 1, 1999; 274(40): 28466 - 28475. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Dye, J. Lee, R. Atkinson, R Brewster, P. Han, and H. Bellen The Drosophila sanpodo gene controls sibling cell fate and encodes a tropomodulin homolog, an actin/tropomyosin-associated protein Development, January 5, 1998; 125(10): 1845 - 1856. [Abstract] [PDF]
|
|
|
|
|
|
A. S. McElhinny, B. Kolmerer, V. M. Fowler, S. Labeit, and C. C. Gregorio The N-terminal End of Nebulin Interacts with Tropomodulin at the Pointed Ends of the Thin Filaments J. Biol. Chem., January 5, 2001; 276(1): 583 - 592. [Abstract] [Full Text] [PDF]
|
|
