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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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Interactions of mBPAG-C1 with aggregates of NF-L/ NF-M proteins. SW13.cl.2Vim− cells from transient transfections  of pcDNA-FLAG-mBPAG-C1 with (A and B) pRSVi-NFL1-415 and pRSVi-NFM; (C and D) pRSVi-NFL and pRSVi-NFM1-421; (E and F) pRSVi-NFL1-415 and pRSVi-NFM1-421 were  double-labeled with mouse anti-FLAG M2 mAb (A, C, and E)  and rabbit polyclonal antibody against NF-L (B, D, and F). A  normal filament network could be formed by wild-type NF-L and  tailless NF-M or by tailless NF-L and wild-type NF-M, but  mBPAG-C1 did not colocalize with these filament networks.  mBPAG-C1 only associated with the aggregates formed by tailless NF-L and tailless NF-M. Bar, 10 μm.
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Figure 7: Interactions of mBPAG-C1 with aggregates of NF-L/ NF-M proteins. SW13.cl.2Vim− cells from transient transfections of pcDNA-FLAG-mBPAG-C1 with (A and B) pRSVi-NFL1-415 and pRSVi-NFM; (C and D) pRSVi-NFL and pRSVi-NFM1-421; (E and F) pRSVi-NFL1-415 and pRSVi-NFM1-421 were double-labeled with mouse anti-FLAG M2 mAb (A, C, and E) and rabbit polyclonal antibody against NF-L (B, D, and F). A normal filament network could be formed by wild-type NF-L and tailless NF-M or by tailless NF-L and wild-type NF-M, but mBPAG-C1 did not colocalize with these filament networks. mBPAG-C1 only associated with the aggregates formed by tailless NF-L and tailless NF-M. Bar, 10 μm.

Mentions: To determine if the tail domains of NFTPs also interfered with the association of mBPAG-C1 with filaments formed by NF-L/NF-M or NF-L/NF-H in the transient transfection assays, we performed cotransfections of mBPAG-C1 with various combinations of tailless and full-length NFTPs. As described previously, normal filament networks are observed from coassembly of tailless NF-L and wild-type NF-M or coassembly of wild-type NF-L and tailless NF-M in transiently transfected SW13.cl.2Vim− cells (Ching and Liem, 1993). However, mBPAG-C1 did not associate with any of the filamentous networks formed from these coassemblies and maintained a diffuse staining pattern (Fig. 7, A–D). mBPAG-C1 only colocalized with aggregates formed by coassembly of tailless NF-L and tailless NF-M (Fig. 7, E and F). These results implied that the tail domains of NF-L and NF-M interfered with the binding of mBPAG-C1 to the filament networks formed by these proteins. When we transfected wild-type NF-H with tailless NF-L in SW13.cl.2Vim− cells, we observed filamentous staining as reported previously (Ching and Liem, 1993). However, mBPAG-C1 showed a diffuse staining pattern without any colocalization with the filaments formed by the tailless NF-L and wild-type NF-H (Fig. 8, A and B). Tailless NF-H formed filaments with wild-type NF-L (Sun et al., 1997), but the coexpressed mBPAG-C1 did not associate with this filament network (Fig. 8, C and D). In contrast, mBPAG-C1 was able to associate with the aggregates formed by tailless NF-L and tailless NF-H (Fig. 8, E and F). These experiments showed that the presence of any of the NFTP tail domains was sufficient to prevent NFTPs from interacting with mBPAG-C1.


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)

Interactions of mBPAG-C1 with aggregates of NF-L/ NF-M proteins. SW13.cl.2Vim− cells from transient transfections  of pcDNA-FLAG-mBPAG-C1 with (A and B) pRSVi-NFL1-415 and pRSVi-NFM; (C and D) pRSVi-NFL and pRSVi-NFM1-421; (E and F) pRSVi-NFL1-415 and pRSVi-NFM1-421 were  double-labeled with mouse anti-FLAG M2 mAb (A, C, and E)  and rabbit polyclonal antibody against NF-L (B, D, and F). A  normal filament network could be formed by wild-type NF-L and  tailless NF-M or by tailless NF-L and wild-type NF-M, but  mBPAG-C1 did not colocalize with these filament networks.  mBPAG-C1 only associated with the aggregates formed by tailless NF-L and tailless NF-M. Bar, 10 μm.
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Figure 7: Interactions of mBPAG-C1 with aggregates of NF-L/ NF-M proteins. SW13.cl.2Vim− cells from transient transfections of pcDNA-FLAG-mBPAG-C1 with (A and B) pRSVi-NFL1-415 and pRSVi-NFM; (C and D) pRSVi-NFL and pRSVi-NFM1-421; (E and F) pRSVi-NFL1-415 and pRSVi-NFM1-421 were double-labeled with mouse anti-FLAG M2 mAb (A, C, and E) and rabbit polyclonal antibody against NF-L (B, D, and F). A normal filament network could be formed by wild-type NF-L and tailless NF-M or by tailless NF-L and wild-type NF-M, but mBPAG-C1 did not colocalize with these filament networks. mBPAG-C1 only associated with the aggregates formed by tailless NF-L and tailless NF-M. Bar, 10 μm.
Mentions: To determine if the tail domains of NFTPs also interfered with the association of mBPAG-C1 with filaments formed by NF-L/NF-M or NF-L/NF-H in the transient transfection assays, we performed cotransfections of mBPAG-C1 with various combinations of tailless and full-length NFTPs. As described previously, normal filament networks are observed from coassembly of tailless NF-L and wild-type NF-M or coassembly of wild-type NF-L and tailless NF-M in transiently transfected SW13.cl.2Vim− cells (Ching and Liem, 1993). However, mBPAG-C1 did not associate with any of the filamentous networks formed from these coassemblies and maintained a diffuse staining pattern (Fig. 7, A–D). mBPAG-C1 only colocalized with aggregates formed by coassembly of tailless NF-L and tailless NF-M (Fig. 7, E and F). These results implied that the tail domains of NF-L and NF-M interfered with the binding of mBPAG-C1 to the filament networks formed by these proteins. When we transfected wild-type NF-H with tailless NF-L in SW13.cl.2Vim− cells, we observed filamentous staining as reported previously (Ching and Liem, 1993). However, mBPAG-C1 showed a diffuse staining pattern without any colocalization with the filaments formed by the tailless NF-L and wild-type NF-H (Fig. 8, A and B). Tailless NF-H formed filaments with wild-type NF-L (Sun et al., 1997), but the coexpressed mBPAG-C1 did not associate with this filament network (Fig. 8, C and D). In contrast, mBPAG-C1 was able to associate with the aggregates formed by tailless NF-L and tailless NF-H (Fig. 8, E and F). These experiments showed that the presence of any of the NFTP tail domains was sufficient to prevent NFTPs from interacting with mBPAG-C1.

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