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The intermediate filament protein peripherin is the specific interaction partner of mouse BPAG1-n (dystonin) in neurons.

Leung CL, Sun D, Liem RK - J. Cell Biol. (1999)

Bottom Line: Kouklis, D.W.Fuchs. 1996.Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

View Article: PubMed Central - PubMed

Affiliation: Departments of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York 10032, USA.

ABSTRACT
The dystonia musculorum (dt) mouse suffers from severe degeneration of primary sensory neurons. The mutated gene product is named dystonin and is identical to the neuronal isoform of bullous pemphigoid antigen 1 (BPAG1-n). BPAG1-n contains an actin-binding domain at its NH2 terminus and a putative intermediate filament-binding domain at its COOH terminus. Because the degenerating sensory neurons of dt mice display abnormal accumulations of intermediate filaments in the axons, BPAG1-n has been postulated to organize the neuronal cytoskeleton by interacting with both the neurofilament triplet proteins (NFTPs) and microfilaments. In this paper we show by a variety of methods that the COOH-terminal tail domain of mouse BPAG1 interacts specifically with peripherin, but in contrast to a previous study (Yang, Y., J. Dowling, Q.C. Yu, P. Kouklis, D.W. Cleveland, and E. Fuchs. 1996. Cell. 86:655-665), mouse BPAG1 fails to associate with full-length NFTPs. The tail domains interfered with the association of the NFTPs with BPAG1. In dt mice, peripherin is present in axonal swellings of degenerating sensory neurons in the dorsal root ganglia and is downregulated even in other neural regions, which have no obvious signs of pathology. Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

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mBPAG-C2 associated with peripherin filaments but  not with NF-L/NF-H filaments in transiently transfected cells.  Transient transfections of pcDNA-FLAG-mBPAG-C2 with  pRSVi-peripherin (A and B) or with pCI-NFL/NFH (C and D)  were performed in SW13.cl.2Vim− cells. Transfected cells were  stained with mouse anti-FLAG M2 mAb (A and C), rabbit polyclonal antibodies against peripherin (B), and rabbit polyclonal  antibody against NF-L (D), and were examined by confocal microscopy. Similar to mBPAG-C1, mBPAG-C2 only colocalized  with peripherin filaments and showed no correlation with the  NF-L/NF-H filaments. Bar, 10 μm.
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Figure 4: mBPAG-C2 associated with peripherin filaments but not with NF-L/NF-H filaments in transiently transfected cells. Transient transfections of pcDNA-FLAG-mBPAG-C2 with pRSVi-peripherin (A and B) or with pCI-NFL/NFH (C and D) were performed in SW13.cl.2Vim− cells. Transfected cells were stained with mouse anti-FLAG M2 mAb (A and C), rabbit polyclonal antibodies against peripherin (B), and rabbit polyclonal antibody against NF-L (D), and were examined by confocal microscopy. Similar to mBPAG-C1, mBPAG-C2 only colocalized with peripherin filaments and showed no correlation with the NF-L/NF-H filaments. Bar, 10 μm.

Mentions: The discrepancy between our results and Yang et al. (1996) could be due to the presence of the partial rod domain in our mBPAG-C1 protein. To investigate this possibility, transient transfections with pcDNA-FLAG-mBPAG-C2 construct were carried out. mBPAG-C2 contains only the mouse BPAG1 tail domain (Fig. 1) and is therefore the mouse equivalent of the human BPAG1 tail domain used by Yang et al. (1996). As shown in Fig. 4 by confocal microscopy, mBPAG-C2 also colocalized with the peripherin filament network, but not with the NF-L/ NF-H filament network. Moreover, mBPAG-C2 disrupted the α-internexin filament network and showed no correlation with the NF-L/NF-M filament network (data not shown). Because identical results were obtained by using either mBPAG-C1 or mBPAG-C2, we concluded that the presence of the partial rod domain in mBPAG-C1 did not affect its interaction with IF networks in transient transfection assays.


The intermediate filament protein peripherin is the specific interaction partner of mouse BPAG1-n (dystonin) in neurons.

Leung CL, Sun D, Liem RK - J. Cell Biol. (1999)

mBPAG-C2 associated with peripherin filaments but  not with NF-L/NF-H filaments in transiently transfected cells.  Transient transfections of pcDNA-FLAG-mBPAG-C2 with  pRSVi-peripherin (A and B) or with pCI-NFL/NFH (C and D)  were performed in SW13.cl.2Vim− cells. Transfected cells were  stained with mouse anti-FLAG M2 mAb (A and C), rabbit polyclonal antibodies against peripherin (B), and rabbit polyclonal  antibody against NF-L (D), and were examined by confocal microscopy. Similar to mBPAG-C1, mBPAG-C2 only colocalized  with peripherin filaments and showed no correlation with the  NF-L/NF-H filaments. Bar, 10 μm.
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Related In: Results  -  Collection

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Figure 4: mBPAG-C2 associated with peripherin filaments but not with NF-L/NF-H filaments in transiently transfected cells. Transient transfections of pcDNA-FLAG-mBPAG-C2 with pRSVi-peripherin (A and B) or with pCI-NFL/NFH (C and D) were performed in SW13.cl.2Vim− cells. Transfected cells were stained with mouse anti-FLAG M2 mAb (A and C), rabbit polyclonal antibodies against peripherin (B), and rabbit polyclonal antibody against NF-L (D), and were examined by confocal microscopy. Similar to mBPAG-C1, mBPAG-C2 only colocalized with peripherin filaments and showed no correlation with the NF-L/NF-H filaments. Bar, 10 μm.
Mentions: The discrepancy between our results and Yang et al. (1996) could be due to the presence of the partial rod domain in our mBPAG-C1 protein. To investigate this possibility, transient transfections with pcDNA-FLAG-mBPAG-C2 construct were carried out. mBPAG-C2 contains only the mouse BPAG1 tail domain (Fig. 1) and is therefore the mouse equivalent of the human BPAG1 tail domain used by Yang et al. (1996). As shown in Fig. 4 by confocal microscopy, mBPAG-C2 also colocalized with the peripherin filament network, but not with the NF-L/ NF-H filament network. Moreover, mBPAG-C2 disrupted the α-internexin filament network and showed no correlation with the NF-L/NF-M filament network (data not shown). Because identical results were obtained by using either mBPAG-C1 or mBPAG-C2, we concluded that the presence of the partial rod domain in mBPAG-C1 did not affect its interaction with IF networks in transient transfection assays.

Bottom Line: Kouklis, D.W.Fuchs. 1996.Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

View Article: PubMed Central - PubMed

Affiliation: Departments of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York 10032, USA.

ABSTRACT
The dystonia musculorum (dt) mouse suffers from severe degeneration of primary sensory neurons. The mutated gene product is named dystonin and is identical to the neuronal isoform of bullous pemphigoid antigen 1 (BPAG1-n). BPAG1-n contains an actin-binding domain at its NH2 terminus and a putative intermediate filament-binding domain at its COOH terminus. Because the degenerating sensory neurons of dt mice display abnormal accumulations of intermediate filaments in the axons, BPAG1-n has been postulated to organize the neuronal cytoskeleton by interacting with both the neurofilament triplet proteins (NFTPs) and microfilaments. In this paper we show by a variety of methods that the COOH-terminal tail domain of mouse BPAG1 interacts specifically with peripherin, but in contrast to a previous study (Yang, Y., J. Dowling, Q.C. Yu, P. Kouklis, D.W. Cleveland, and E. Fuchs. 1996. Cell. 86:655-665), mouse BPAG1 fails to associate with full-length NFTPs. The tail domains interfered with the association of the NFTPs with BPAG1. In dt mice, peripherin is present in axonal swellings of degenerating sensory neurons in the dorsal root ganglia and is downregulated even in other neural regions, which have no obvious signs of pathology. Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

Show MeSH
Related in: MedlinePlus