Emerin binding to Btf, a death-promoting transcriptional repressor, is disrupted by a missense mutation that causes Emery-Dreifuss muscular dystrophy

Tokuko Haraguchi, James M. Holaska, Miho Yamane, Takako Koujin, Noriyo Hashiguchi, Chie Mori, Katherine Lee Wilson, Yasushi Hiraoka

Research output: Contribution to journalArticle

Abstract

Loss of functional emerin, a nuclear membrane protein, causes X-linked recessive Emery-Dreifuss muscular dystrophy. In a yeast two-hybrid screen, we found that emerin interacts with Btf, a death-promoting transcriptional repressor, which is expressed at high levels in skeletal muscle. Biochemical analysis showed that emerin binds Btf with an equilibrium affinity (K D) of 100 nM. Using a collection of 21 clustered alanine- substitution mutations in emerin, the residues required for binding to Btf mapped to two regions of emerin that flank its lamin-binding domain. Two disease-causing mutations in emerin, S54F and Δ95-99, disrupted binding to Btf. The Δ95-99 mutation was relatively uninformative, as this mutation also disrupts emerin binding to lamin A and a different transcription repressor named germ cell-less (GCL). In striking contrast, emerin mutant S54F, which binds normally to barrier-to-autointegration factor, lamin A and GCL, selectively disrupted emerin binding to Btf. We localized endogenous Btf in HeLa cells by indirect immunoflurorescence using affinity-purified antibodies against Btf. In nonapoptotic HeLa cells Btf was found in dot-like structures throughout the nuclear interior. However, within 3 h after treating cells with Fas antibody to induce apoptosis, the distribution of Btf changed, and Btf concentrated in a distinct zone near the nuclear envelope. These results suggest that Btf localization is regulated by apoptotic signals, and that loss of emerin binding to Btf may be relevant to muscle wasting in Emery-Dreifuss muscular dystrophy.

Original languageEnglish (US)
Pages (from-to)1035-1045
Number of pages11
JournalEuropean Journal of Biochemistry
Volume271
Issue number5
DOIs
StatePublished - Mar 2004

Fingerprint

Emery-Dreifuss Muscular Dystrophy
Missense Mutation
Lamin Type A
Mutation
Nuclear Envelope
HeLa Cells
Germ Cells
Muscle
emerin
Lamins
Antibody Affinity
Antibodies
Transcription
Nuclear Proteins
Alanine
Yeast
Membrane Proteins
Skeletal Muscle
Substitution reactions

Keywords

  • Apoptosis
  • Emerin
  • Emery-Dreifuss muscular dystrophy
  • Lamin A
  • MAN1

ASJC Scopus subject areas

  • Biochemistry

Cite this

Emerin binding to Btf, a death-promoting transcriptional repressor, is disrupted by a missense mutation that causes Emery-Dreifuss muscular dystrophy. / Haraguchi, Tokuko; Holaska, James M.; Yamane, Miho; Koujin, Takako; Hashiguchi, Noriyo; Mori, Chie; Wilson, Katherine Lee; Hiraoka, Yasushi.

In: European Journal of Biochemistry, Vol. 271, No. 5, 03.2004, p. 1035-1045.

Research output: Contribution to journalArticle

Haraguchi, Tokuko ; Holaska, James M. ; Yamane, Miho ; Koujin, Takako ; Hashiguchi, Noriyo ; Mori, Chie ; Wilson, Katherine Lee ; Hiraoka, Yasushi. / Emerin binding to Btf, a death-promoting transcriptional repressor, is disrupted by a missense mutation that causes Emery-Dreifuss muscular dystrophy. In: European Journal of Biochemistry. 2004 ; Vol. 271, No. 5. pp. 1035-1045.
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abstract = "Loss of functional emerin, a nuclear membrane protein, causes X-linked recessive Emery-Dreifuss muscular dystrophy. In a yeast two-hybrid screen, we found that emerin interacts with Btf, a death-promoting transcriptional repressor, which is expressed at high levels in skeletal muscle. Biochemical analysis showed that emerin binds Btf with an equilibrium affinity (K D) of 100 nM. Using a collection of 21 clustered alanine- substitution mutations in emerin, the residues required for binding to Btf mapped to two regions of emerin that flank its lamin-binding domain. Two disease-causing mutations in emerin, S54F and Δ95-99, disrupted binding to Btf. The Δ95-99 mutation was relatively uninformative, as this mutation also disrupts emerin binding to lamin A and a different transcription repressor named germ cell-less (GCL). In striking contrast, emerin mutant S54F, which binds normally to barrier-to-autointegration factor, lamin A and GCL, selectively disrupted emerin binding to Btf. We localized endogenous Btf in HeLa cells by indirect immunoflurorescence using affinity-purified antibodies against Btf. In nonapoptotic HeLa cells Btf was found in dot-like structures throughout the nuclear interior. However, within 3 h after treating cells with Fas antibody to induce apoptosis, the distribution of Btf changed, and Btf concentrated in a distinct zone near the nuclear envelope. These results suggest that Btf localization is regulated by apoptotic signals, and that loss of emerin binding to Btf may be relevant to muscle wasting in Emery-Dreifuss muscular dystrophy.",
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AU - Holaska, James M.

AU - Yamane, Miho

AU - Koujin, Takako

AU - Hashiguchi, Noriyo

AU - Mori, Chie

AU - Wilson, Katherine Lee

AU - Hiraoka, Yasushi

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AB - Loss of functional emerin, a nuclear membrane protein, causes X-linked recessive Emery-Dreifuss muscular dystrophy. In a yeast two-hybrid screen, we found that emerin interacts with Btf, a death-promoting transcriptional repressor, which is expressed at high levels in skeletal muscle. Biochemical analysis showed that emerin binds Btf with an equilibrium affinity (K D) of 100 nM. Using a collection of 21 clustered alanine- substitution mutations in emerin, the residues required for binding to Btf mapped to two regions of emerin that flank its lamin-binding domain. Two disease-causing mutations in emerin, S54F and Δ95-99, disrupted binding to Btf. The Δ95-99 mutation was relatively uninformative, as this mutation also disrupts emerin binding to lamin A and a different transcription repressor named germ cell-less (GCL). In striking contrast, emerin mutant S54F, which binds normally to barrier-to-autointegration factor, lamin A and GCL, selectively disrupted emerin binding to Btf. We localized endogenous Btf in HeLa cells by indirect immunoflurorescence using affinity-purified antibodies against Btf. In nonapoptotic HeLa cells Btf was found in dot-like structures throughout the nuclear interior. However, within 3 h after treating cells with Fas antibody to induce apoptosis, the distribution of Btf changed, and Btf concentrated in a distinct zone near the nuclear envelope. These results suggest that Btf localization is regulated by apoptotic signals, and that loss of emerin binding to Btf may be relevant to muscle wasting in Emery-Dreifuss muscular dystrophy.

KW - Apoptosis

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