LAP2 binds to BAF•DNA complexes:: requirement for the LEM domain and modulation by variable regions

被引:172
作者
Shumaker, DK
Lee, KK
Tanhehco, YC
Craigie, R
Wilson, KL
机构
[1] Johns Hopkins Univ, Sch Med, Dept Cell Biol & Anat, Baltimore, MD 21205 USA
[2] NIDDK, Mol Biol Lab, NIH, Bethesda, MD 20892 USA
关键词
barrier-to-autointegration factor; emerin; Emery-Dreifuss muscular dystrophy; lamin-associated polypeptide 2 (LAP2); nuclear envelope;
D O I
10.1093/emboj/20.7.1754
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
LAP2 belongs to a family of nuclear membrane proteins sharing a 43 residue LEM domain. All LAP2 isoforms have the same N-terminal 'constant' region (LAP2-c), which includes the LEM domain, plus a C-terminal 'variable' region. LAP2-c polypeptide inhibits nuclear assembly in Xenopus extracts, and binds in vitro to barrier-to-autointegration factor (BAF), a DNA-bridging protein. We tested 17 Xenopus LAP2-c mutants for nuclear assembly inhibition, and binding to BAF and BAB DNA complexes. LEM domain mutations disrupted all activities tested. Some mutations outside the LEM domain had no effect on binding to BAF, but disrupted activity in Xenopus extracts, suggesting that LAP2-c has an additional unknown function required to inhibit nuclear assembly. Mutagenesis results suggest that BAF changes conformation when complexed with DNA. The binding affinity of LAP2 was higher for BAF DNA complexes than for BAF, suggesting that these interactions are physiologically relevant. Nucleoplasmic domains of Xenopus LAP2 isoforms varied 9-fold in their affinities for BAF, but all isoforms supershifted BAP DNA complexes. We propose that the LEM domain is a core BAF-binding domain that can be modulated by the variable regions of LAP2 isoforms.
引用
收藏
页码:1754 / 1764
页数:11
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