Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.

The translocation of macromolecules into the nucleus is a fundamental eukaryotic process, regulating gene expression, cell division and differentiation, but which is impaired in a range of significant diseases including cancer and viral infection. The import of proteins into the nucleus is generally...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Noelia Roman, Mary Christie, Crystall M D Swarbrick, Bostjan Kobe, Jade K Forwood
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3029e82fd2e9428a8402e883fde5c545
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:3029e82fd2e9428a8402e883fde5c545
record_format	dspace
spelling	oai:doaj.org-article:3029e82fd2e9428a8402e883fde5c5452021-11-18T08:42:42ZStructural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.1932-620310.1371/journal.pone.0082038https://doaj.org/article/3029e82fd2e9428a8402e883fde5c5452013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24339986/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The translocation of macromolecules into the nucleus is a fundamental eukaryotic process, regulating gene expression, cell division and differentiation, but which is impaired in a range of significant diseases including cancer and viral infection. The import of proteins into the nucleus is generally initiated by a specific, high affinity interaction between nuclear localisation signals (NLSs) and nuclear import receptors in the cytoplasm, and terminated through the disassembly of these complexes in the nucleus. For classical NLSs (cNLSs), this import is mediated by the importin-α (IMPα) adaptor protein, which in turn binds to IMPβ to mediate translocation of nuclear cargo across the nuclear envelope. The interaction and disassembly of import receptor:cargo complexes is reliant on the differential localisation of nucleotide bound Ran across the envelope, maintained in its low affinity, GDP-bound form in the cytoplasm, and its high affinity, GTP-bound form in the nucleus. This in turn is maintained by the differential localisation of Ran regulating proteins, with RanGAP in the cytoplasm maintaining Ran in its GDP-bound form, and RanGEF (Prp20 in yeast) in the nucleus maintaining Ran in its GTP-bound form. Here, we describe the 2.1 Å resolution x-ray crystal structure of IMPα in complex with the NLS of Prp20. We observe 1,091 Å(2) of buried surface area mediated by an extensive array of contacts involving residues on armadillo repeats 2-7, utilising both the major and minor NLS binding sites of IMPα to contact bipartite NLS clusters (17)RAKKMSK(23) and (3)KR(4), respectively. One notable feature of the major site is the insertion of Prp20NLS Ala(18) between the P0 and P1 NLS sites, noted in only a few classical bipartite NLSs. This study provides a detailed account of the binding mechanism enabling Prp20 interaction with the nuclear import receptor, and additional new information for the interaction between IMPα and cargo.Noelia RomanMary ChristieCrystall M D SwarbrickBostjan KobeJade K ForwoodPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 12, p e82038 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Noelia Roman Mary Christie Crystall M D Swarbrick Bostjan Kobe Jade K Forwood Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.
description	The translocation of macromolecules into the nucleus is a fundamental eukaryotic process, regulating gene expression, cell division and differentiation, but which is impaired in a range of significant diseases including cancer and viral infection. The import of proteins into the nucleus is generally initiated by a specific, high affinity interaction between nuclear localisation signals (NLSs) and nuclear import receptors in the cytoplasm, and terminated through the disassembly of these complexes in the nucleus. For classical NLSs (cNLSs), this import is mediated by the importin-α (IMPα) adaptor protein, which in turn binds to IMPβ to mediate translocation of nuclear cargo across the nuclear envelope. The interaction and disassembly of import receptor:cargo complexes is reliant on the differential localisation of nucleotide bound Ran across the envelope, maintained in its low affinity, GDP-bound form in the cytoplasm, and its high affinity, GTP-bound form in the nucleus. This in turn is maintained by the differential localisation of Ran regulating proteins, with RanGAP in the cytoplasm maintaining Ran in its GDP-bound form, and RanGEF (Prp20 in yeast) in the nucleus maintaining Ran in its GTP-bound form. Here, we describe the 2.1 Å resolution x-ray crystal structure of IMPα in complex with the NLS of Prp20. We observe 1,091 Å(2) of buried surface area mediated by an extensive array of contacts involving residues on armadillo repeats 2-7, utilising both the major and minor NLS binding sites of IMPα to contact bipartite NLS clusters (17)RAKKMSK(23) and (3)KR(4), respectively. One notable feature of the major site is the insertion of Prp20NLS Ala(18) between the P0 and P1 NLS sites, noted in only a few classical bipartite NLSs. This study provides a detailed account of the binding mechanism enabling Prp20 interaction with the nuclear import receptor, and additional new information for the interaction between IMPα and cargo.
format	article
author	Noelia Roman Mary Christie Crystall M D Swarbrick Bostjan Kobe Jade K Forwood
author_facet	Noelia Roman Mary Christie Crystall M D Swarbrick Bostjan Kobe Jade K Forwood
author_sort	Noelia Roman
title	Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.
title_short	Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.
title_full	Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.
title_fullStr	Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.
title_full_unstemmed	Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.
title_sort	structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite nls of prp20.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/3029e82fd2e9428a8402e883fde5c545
work_keys_str_mv	AT noeliaroman structuralcharacterisationofthenuclearimportreceptorimportinalphaincomplexwiththebipartitenlsofprp20 AT marychristie structuralcharacterisationofthenuclearimportreceptorimportinalphaincomplexwiththebipartitenlsofprp20 AT crystallmdswarbrick structuralcharacterisationofthenuclearimportreceptorimportinalphaincomplexwiththebipartitenlsofprp20 AT bostjankobe structuralcharacterisationofthenuclearimportreceptorimportinalphaincomplexwiththebipartitenlsofprp20 AT jadekforwood structuralcharacterisationofthenuclearimportreceptorimportinalphaincomplexwiththebipartitenlsofprp20
_version_	1718421373121986560

Structural characterisation of the nuclear import receptor importin alpha in complex with the bipartite NLS of Prp20.

Ejemplares similares