Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)

Abstract Mutations in the RNA-binding protein FUS cause familial amyotropic lateral sclerosis (ALS). Several mutations that affect the proline-tyrosine nuclear localization signal (PY-NLS) of FUS cause severe juvenile ALS. FUS also undergoes liquid–liquid phase separation (LLPS) to accumulate in str...

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Autores principales:	Abner Gonzalez, Taro Mannen, Tolga Çağatay, Ayano Fujiwara, Hiroyoshi Matsumura, Ashley B. Niesman, Chad A. Brautigam, Yuh Min Chook, Takuya Yoshizawa
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b02a0e699ad84a3cbf771fac666ddd4f
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spelling	oai:doaj.org-article:b02a0e699ad84a3cbf771fac666ddd4f2021-12-02T14:26:54ZMechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)10.1038/s41598-021-83196-y2045-2322https://doaj.org/article/b02a0e699ad84a3cbf771fac666ddd4f2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83196-yhttps://doaj.org/toc/2045-2322Abstract Mutations in the RNA-binding protein FUS cause familial amyotropic lateral sclerosis (ALS). Several mutations that affect the proline-tyrosine nuclear localization signal (PY-NLS) of FUS cause severe juvenile ALS. FUS also undergoes liquid–liquid phase separation (LLPS) to accumulate in stress granules when cells are stressed. In unstressed cells, wild type FUS resides predominantly in the nucleus as it is imported by the importin Karyopherin-β2 (Kapβ2), which binds with high affinity to the C-terminal PY-NLS of FUS. Here, we analyze the interactions between two ALS-related variants FUS(P525L) and FUS(R495X) with importins, especially Kapβ2, since they are still partially localized to the nucleus despite their defective/missing PY-NLSs. The crystal structure of the Kapβ2·FUS(P525L)PY-NLS complex shows the mutant peptide making fewer contacts at the mutation site, explaining decreased affinity for Kapβ2. Biochemical analysis revealed that the truncated FUS(R495X) protein, although missing the PY-NLS, can still bind Kapβ2 and suppresses LLPS. FUS(R495X) uses its C-terminal tandem arginine-glycine-glycine regions, RGG2 and RGG3, to bind the PY-NLS binding site of Kapβ2 for nuclear localization in cells when arginine methylation is inhibited. These findings suggest the importance of the C-terminal RGG regions in nuclear import and LLPS regulation of ALS variants of FUS that carry defective PY-NLSs.Abner GonzalezTaro MannenTolga ÇağatayAyano FujiwaraHiroyoshi MatsumuraAshley B. NiesmanChad A. BrautigamYuh Min ChookTakuya YoshizawaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Abner Gonzalez Taro Mannen Tolga Çağatay Ayano Fujiwara Hiroyoshi Matsumura Ashley B. Niesman Chad A. Brautigam Yuh Min Chook Takuya Yoshizawa Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)
description	Abstract Mutations in the RNA-binding protein FUS cause familial amyotropic lateral sclerosis (ALS). Several mutations that affect the proline-tyrosine nuclear localization signal (PY-NLS) of FUS cause severe juvenile ALS. FUS also undergoes liquid–liquid phase separation (LLPS) to accumulate in stress granules when cells are stressed. In unstressed cells, wild type FUS resides predominantly in the nucleus as it is imported by the importin Karyopherin-β2 (Kapβ2), which binds with high affinity to the C-terminal PY-NLS of FUS. Here, we analyze the interactions between two ALS-related variants FUS(P525L) and FUS(R495X) with importins, especially Kapβ2, since they are still partially localized to the nucleus despite their defective/missing PY-NLSs. The crystal structure of the Kapβ2·FUS(P525L)PY-NLS complex shows the mutant peptide making fewer contacts at the mutation site, explaining decreased affinity for Kapβ2. Biochemical analysis revealed that the truncated FUS(R495X) protein, although missing the PY-NLS, can still bind Kapβ2 and suppresses LLPS. FUS(R495X) uses its C-terminal tandem arginine-glycine-glycine regions, RGG2 and RGG3, to bind the PY-NLS binding site of Kapβ2 for nuclear localization in cells when arginine methylation is inhibited. These findings suggest the importance of the C-terminal RGG regions in nuclear import and LLPS regulation of ALS variants of FUS that carry defective PY-NLSs.
format	article
author	Abner Gonzalez Taro Mannen Tolga Çağatay Ayano Fujiwara Hiroyoshi Matsumura Ashley B. Niesman Chad A. Brautigam Yuh Min Chook Takuya Yoshizawa
author_facet	Abner Gonzalez Taro Mannen Tolga Çağatay Ayano Fujiwara Hiroyoshi Matsumura Ashley B. Niesman Chad A. Brautigam Yuh Min Chook Takuya Yoshizawa
author_sort	Abner Gonzalez
title	Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)
title_short	Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)
title_full	Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)
title_fullStr	Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)
title_full_unstemmed	Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)
title_sort	mechanism of karyopherin-β2 binding and nuclear import of als variants fus(p525l) and fus(r495x)
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b02a0e699ad84a3cbf771fac666ddd4f
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Mechanism of karyopherin-β2 binding and nuclear import of ALS variants FUS(P525L) and FUS(R495X)

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