© 1995 Oxford University Press
RESEARCH-ARTICLE |
Cloning and characterization of alternatively spliced isoforms of Dp71
Department of Pathology, McMaster University and the Hamilton Civic Hospitals Research Centre Hamilton, Ontario, L8V 1C3 1Genetics Department and Research Institute, The Hospital for Sick Children Children, 555 University Avenue, Toronto, Ontario, M5G 1X8 2The Ontario Cancer Institute 500 Sherbourne Street, Toronto, Ontario, Canada, M4X 1K9
*To whom correspondence should be addressed
Received March 16, 1995; Accepted May 29, 1995
Dp71, a C-terminal isoform of dystrophin, has been identified as the major DMD gene product in many nonmuscle tissues. In this report, reverse transcriptase-polymerase chain reaction (RT-PCR) was used to clone and characterize four alternatively spliced Dp71 transcripts from cultured human amniocytes. The cDNAs encoding these Dp71 transcripts were shown to be alternatively spliced for exons 71 and/or 78. RT-PCR anawlysis also revealed that Dp71 transcripts alternatively spliced for exons 71 and/or 78 were expressed at varying levels in a number of adult human tissues, including muscle, heart, brain, kidney, lung, testis and liver. To investigate size heterogeneity at the translational level, Dp71 cDNAs isolated from amniocytes were expressed in E.coli to generate recombinant Dp71 fusion proteins. These fusion proteins were identified on immunoblots using antibodies specific for the C-terminal sequences of dystrophin that either included (antibody 1461) or excluded exon 78 (antibody 462B). The molecular masses of the Dp71 fusion proteins ranged from 71–75 kDa on SDS-PAGE, consistent with their predicted values. Immunoblot analysis using antibodies 1461 and 462B identified multiple Dp71 isoforms of approximately 70–75 kDa on SDS-PAGE in total protein lysates from amniocytes and various adult human tissues. This variation in molecular mass is consistent with the expression of Dp71 isoforms derived from transcripts alternatively spliced for exons 71 and/or 78. Total protein lysates from normal skeletal muscle, DMD muscle, amniocytes and brain were shown to contain ß-dystroglycan, a component of the dystrophin-associated glycoprotein complex (DGC). Taken together, these results indicate that Dp71 is alternatively spliced for exons 71 and/or 78, and encodes multiple protein isoforms in a wide range of human tissues. The co-expression of these isoforms with ß-dystroglycan suggests that they may form part of a ‘nonmuscle’ DGC somewhat similar to that observed in muscle.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  T. Yokota, Q.-L. Lu, J. E. Morgan, K. E. Davies, R. Fisher, S. Takeda, and T. A. Partridge Expansion of revertant fibers in dystrophic mdx muscles reflects activity of muscle precursor cells and serves as an index of muscle regeneration J. Cell Sci., July 1, 2006; 119(13): 2679 - 2687. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 U. Pozzoli, G. Elgar, R. Cagliani, L. Riva, G. P. Comi, N. Bresolin, A. Bardoni, and M. Sironi Comparative Analysis of Vertebrate Dystrophin Loci Indicate Intron Gigantism as a Common Feature Genome Res., May 1, 2003; 13(5): 764 - 772. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. C. Austin, J. E. B. Fox, G. H. Werstuck, A. R. Stafford, D. E. Bulman, G. Y. Dally, C. A. Ackerley, J. I. Weitz, and P. N. Ray Identification of Dp71 Isoforms in the Platelet Membrane Cytoskeleton. POTENTIAL ROLE IN THROMBIN-MEDIATED PLATELET ADHESION J. Biol. Chem., November 27, 2002; 277(49): 47106 - 47113. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Yang, S. A. Shaver, A. J. Hilliker, and M. B. Sokolowski Abnormal Turning Behavior in Drosophila Larvae: Identification and Molecular Analysis of scribbler (sbb) Genetics, July 1, 2000; 155(3): 1161 - 1174. [Abstract] [Full Text]
|
|
|
|
|
|
T Claudepierre, C Dalloz, D Mornet, K Matsumura, J Sahel, and A Rendon Characterization of the intermolecular associations of the dystrophin-associated glycoprotein complex in retinal Muller glial cells J. Cell Sci., January 10, 2000; 113(19): 3409 - 3417. [Abstract] [PDF]
|
|
|
|
 |
|
 T. Claudepierre, D. Mornet, T. Pannicke, V. Forster, C. Dalloz, F. Bolaños, J. Sahel, A. Reichenbach, and A. Rendon Expression of Dp71 in Muller Glial Cells: A Comparison with Utrophin- and Dystrophin-Associated Proteins Invest. Ophthalmol. Vis. Sci., January 1, 2000; 41(1): 294 - 304. [Abstract] [Full Text]
|
|
|
|
|
|
P. C. Y. Liaw, J. C. Fredenburgh, A. R. Stafford, A. Tulinsky, R. C. Austin, and J. I. Weitz Localization of the Thrombin-binding Domain on Prothrombin Fragment 2 J. Biol. Chem., April 10, 1998; 273(15): 8932 - 8939. [Abstract] [Full Text] [PDF]
|
|