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Nspl1, a new Z-band-associated protein

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Abstract

Molecular characterization of a novel gene designated Neuroendocrine-Specific Protein-Like-1 (Nspl1) had revealed that this gene is expressed as two transcripts, a 1.2 kb transcript found predominantly in skeletal muscle and a 2.1 kb transcript expressed in the brain. The exceptionally high level of skeletal muscle expression prompted us to determine where the protein is localized to skeletal muscle. In vitro studies were performed using two plasmid constructs that generate full-length Nspl1 muscle-specific protein fused to the green fluorescent protein (GFP). In one construct, the GFP cDNA was fused to the N-terminus of the Nspl1 cDNA while in the second construct, the GFP cDNA was fused to the C-terminus of the Nspl1 cDNA. Transfection of either plasmid into mononucleated myoblasts showed that the Nspl1-GFP chimeric protein was associated with intermediate filaments. This was confirmed by using an antibody to stain desmin and finding that GFP-Nspl1 colocalizes with desmin. Chick primary myoblasts were transfected with the chimeric cDNAs and allowed to differentiate into mature myotubes. Results from this analysis and the use of monoclonal antibody to stain α-actinin, further localized the Nspl1 protein to the Z-band of mature myotubes. Confocal microscopy of the myotubes containing Nspl1-GFP demonstrates that Nspl1 is distributed continuously throughout the Z-disks.

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Authors and Affiliations

  1. University of Tennessee Graduate School of Biomedical Sciences, University of Tennessee, Knoxville, TN, 37932, USA

    John G. Geisler

  2. Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN, 37932, USA

    Robert J. Palmer

  3. Human Genome Center, Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, 7000 East Ave., Livermore, CA, 94550, USA

    Lisa J. Stubbs

  4. Life Sciences Division, Oak Ridge National Laboratory, P.O. Box 2009, Oak Ridge, TN, 37831-8077, USA

    Michael L. Mucenski

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  1. John G. Geisler
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  2. Robert J. Palmer
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  3. Lisa J. Stubbs
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  4. Michael L. Mucenski
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Correspondence to John G. Geisler.

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Geisler, J.G., Palmer, R.J., Stubbs, L.J. et al. Nspl1, a new Z-band-associated protein. J Muscle Res Cell Motil 20, 661–668 (1999). https://doi.org/10.1023/A:1005533013926

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