Structures of the tRNA export factor in the nuclear and cytosolic states.
Transfer RNAs are among the most ubiquitous molecules in cells, central to decoding information from messenger RNAs on translating ribosomes. In eukaryotic cells, tRNAs are actively transported from their site of synthesis in the nucleus to their site of function in the cytosol. This is mediated by a dedicated nucleo-cytoplasmic transport factor of the karyopherin-beta family (Xpot, also known as Los1 in Saccharomyces cerevisiae). Here we report the 3.2 A resolution structure of Schizosaccharomyces pombe Xpot in complex with tRNA and RanGTP, and the 3.1 A structure of unbound Xpot, revealing both nuclear and cytosolic snapshots of this transport factor. Xpot undergoes a large conformational change on binding cargo, wrapping around the tRNA and, in particular, binding to the tRNA 5' and 3' ends. The binding mode explains how Xpot can recognize all mature tRNAs in the cell and yet distinguish them from those that have not been properly processed, thus coupling tRNA export to quality control.
Cook
Atlanta G
AG
Structural Cell Biology, MPI for Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany.
Fukuhara
Noemi
N
Jinek
Martin
M
Conti
Elena
E
D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism*; D018360:Crystallography, X-Ray; D003600:Cytosol / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D034443:RNA Transport; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012360:RNA, Transfer, Phe / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity; D020931:ran GTP-Binding Protein / Q000737:chemistry / Q000378:metabolism
D020690:GTPase-Activating Proteins; D028861:Nuclear Pore Complex Proteins; D012331:RNA, Fungal; D012360:RNA, Transfer, Phe; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; C543934:Xpot protein, S pombe; D012343:RNA, Transfer; D020931:ran GTP-Binding Protein
Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3
15299374
Nat Rev Mol Cell Biol. 2007 Oct;8(10):761-73
17786152
Science. 1998 Dec 11;282(5396):2082-5
9851929
Mol Cell. 2005 Apr 29;18(3):355-67
15866177
Nucleic Acids Res. 2006;34(17):4711-21
16963774
Biol Cell. 2004 Oct;96(8):639-55
15519698
Nature. 1999 May 20;399(6733):230-7
10353245
Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2587-91
8146159
EMBO J. 2002 Nov 15;21(22):6216-24
12426393
Curr Opin Chem Biol. 2008 Apr;12(2):126-33
18294973
Nucleic Acids Res. 1998 Jan 1;26(1):148-53
9399820
Mol Cell Biol. 2002 Aug;22(16):5708-20
12138183
Curr Opin Struct Biol. 2009 Apr;19(2):226-32
19327984
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W615-9
15215462
Biochemistry. 1985 Dec 3;24(25):7263-8
3910099
RNA Biol. 2009 Jan-Mar;6(1):40-2
19106627
Cell Mol Life Sci. 2007 Sep;64(18):2404-12
17599240
FEBS Lett. 1996 Apr 22;384(3):219-21
8617357
RNA. 1999 Apr;5(4):539-49
10199570
EMBO J. 1998 Dec 15;17(24):7430-41
9857198
Mol Cell. 1998 Feb;1(3):359-69
9660920
Histochem Cell Biol. 2008 Feb;129(2):105-16
18228033
Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21
9757107
Cell. 1999 May 28;97(5):635-46
10367892
Acta Crystallogr D Biol Crystallogr. 2002 Oct;58(Pt 10 Pt 2):1772-9
12351820
J Mol Biol. 2001 May 25;309(1):1-18
11491282
Nucleic Acids Res. 2005 Jul 1;33(Web Server issue):W299-302
15980475
Bioessays. 2009 Apr;31(4):466-77
19274657
Chromosoma. 2006 Apr;115(2):98-109
16421734
Nature. 2004 Dec 16;432(7019):872-7
15602554
J Synchrotron Radiat. 2006 Mar;13(Pt 2):120-30
16495612
Biotechnology (N Y). 1994 Jun;12(6):601-5
7764949
Nature. 1999 May 20;399(6733):221-9
10353244
Trends Biochem Sci. 1992 Apr;17(4):159-64
1585461
Annu Rev Biochem. 1987;56:263-87
3304135
Nat Struct Mol Biol. 2009 May;16(5):558-60
19339972
J Biol Chem. 2004 Jan 16;279(3):2176-81
14561738
Nature. 2005 Jun 2;435(7042):693-6
15864302
J Mol Biol. 2000 Aug 11;301(2):401-14
10926517
RNA. 2000 Aug;6(8):1091-105
10943889
Curr Biol. 1998 Mar 12;8(6):305-14
9512417
Proc Natl Acad Sci U S A. 1983 Nov;80(21):6436-40
6579529
Curr Opin Struct Biol. 2006 Feb;16(1):12-7
16364630
J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
19461840
Mol Cell Biol. 1998 Nov;18(11):6374-86
9774653
Annu Rev Biochem. 2007;76:647-71
17506639
Science. 2009 May 22;324(5930):1087-91
19389996