TY - JOUR
T1 - Nesprin-1α self-associates and binds directly to emerin and lamin A in vitro
AU - Mislow, John M.K.
AU - Holaska, James M.
AU - Kim, Marian S.
AU - Lee, Kenneth K.
AU - Segura-Totten, Miriam
AU - Wilson, Katherine L.
AU - McNally, Elizabeth M.
N1 - Funding Information:
This work was supported by NIH HL63783, the Muscular Dystrophy Association, the American Heart Association and the Burroughs Wellcome Fund (E.M.M.) and NIH GM48646 (K.L.W.). J.M.K. is supported by NIH HL-07237 and HD-07009.
PY - 2002/8/14
Y1 - 2002/8/14
N2 - Nesprin-1α is a spectrin repeat (SR)-containing, transmembrane protein of the inner nuclear membrane, and is highly expressed in muscle cells. A yeast two-hybrid screen for nesprin-1α-interacting proteins showed that nesprin-1α interacted with itself. Blot overlay experiments revealed that nesprin-1α's third SR binds the fifth SR. The carboxy-terminal half of nesprin-1α directly bound lamin A, a nuclear intermediate filament protein. Biochemical analysis demonstrated that nesprin-1α dimers bind directly to the nucleoplasmic domain of emerin, an inner nuclear membrane protein, with an affinity of 4 nM. Binding was optimal for full nucleoplasmic dimers of nesprin-1α, since nesprin fragments SR1-5 and SR5-7 bound emerin as monomers with affinities of 53 nM and 250 mM, respectively. We propose that membrane-anchored nesprin-1α antiparallel dimers interact with both emerin and lamin A to provide scaffolding at the inner nuclear membrane.
AB - Nesprin-1α is a spectrin repeat (SR)-containing, transmembrane protein of the inner nuclear membrane, and is highly expressed in muscle cells. A yeast two-hybrid screen for nesprin-1α-interacting proteins showed that nesprin-1α interacted with itself. Blot overlay experiments revealed that nesprin-1α's third SR binds the fifth SR. The carboxy-terminal half of nesprin-1α directly bound lamin A, a nuclear intermediate filament protein. Biochemical analysis demonstrated that nesprin-1α dimers bind directly to the nucleoplasmic domain of emerin, an inner nuclear membrane protein, with an affinity of 4 nM. Binding was optimal for full nucleoplasmic dimers of nesprin-1α, since nesprin fragments SR1-5 and SR5-7 bound emerin as monomers with affinities of 53 nM and 250 mM, respectively. We propose that membrane-anchored nesprin-1α antiparallel dimers interact with both emerin and lamin A to provide scaffolding at the inner nuclear membrane.
UR - http://www.scopus.com/inward/record.url?scp=0037077382&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037077382&partnerID=8YFLogxK
U2 - 10.1016/S0014-5793(02)03105-8
DO - 10.1016/S0014-5793(02)03105-8
M3 - Article
C2 - 12163176
AN - SCOPUS:0037077382
SN - 0014-5793
VL - 525
SP - 135
EP - 140
JO - FEBS Letters
JF - FEBS Letters
IS - 1-3
ER -