Binding stoichiometry and structural model of the HIV-1 Rev/importin β complex.
Journal
Life science alliance
ISSN: 2575-1077
Titre abrégé: Life Sci Alliance
Pays: United States
ID NLM: 101728869
Informations de publication
Date de publication:
10 2022
10 2022
Historique:
received:
28
02
2022
revised:
21
07
2022
accepted:
22
07
2022
entrez:
22
8
2022
pubmed:
23
8
2022
medline:
25
8
2022
Statut:
epublish
Résumé
HIV-1 Rev mediates the nuclear export of intron-containing viral RNA transcripts and is essential for viral replication. Rev is imported into the nucleus by the host protein importin β (Impβ), but how Rev associates with Impβ is poorly understood. Here, we report biochemical, mutational, and biophysical studies of the Impβ/Rev complex. We show that Impβ binds two Rev monomers through independent binding sites, in contrast to the 1:1 binding stoichiometry observed for most Impβ cargos. Peptide scanning data and charge-reversal mutations identify the N-terminal tip of Rev helix α2 within Rev's arginine-rich motif (ARM) as a primary Impβ-binding epitope. Cross-linking mass spectrometry and compensatory mutagenesis data combined with molecular docking simulations suggest a structural model in which one Rev monomer binds to the C-terminal half of Impβ with Rev helix α2 roughly parallel to the HEAT-repeat superhelical axis, whereas the other monomer binds to the N-terminal half. These findings shed light on the molecular basis of Rev recognition by Impβ and highlight an atypical binding behavior that distinguishes Rev from canonical cellular Impβ cargos.
Identifiants
pubmed: 35995566
pii: 5/10/e202201431
doi: 10.26508/lsa.202201431
pmc: PMC9396022
pii:
doi:
Substances chimiques
RNA, Viral
0
beta Karyopherins
0
Banques de données
PDB
['1UKL', '3W5K', '1QGK', '3LWW', '2X7L', '5DHV']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Informations de copyright
© 2022 Spittler et al.
Références
Mol Cell. 2001 Mar;7(3):603-14
pubmed: 11463385
Biophys J. 2013 Aug 20;105(4):1004-17
pubmed: 23972852
Traffic. 2014 Jul;15(7):727-48
pubmed: 24766099
Protein Sci. 2014 Jun;23(6):747-59
pubmed: 24639379
Biochim Biophys Acta. 2011 Sep;1813(9):1593-606
pubmed: 21029754
Biochemistry. 2010 Jun 22;49(24):5042-7
pubmed: 20476751
J Virol. 2017 Jan 18;91(3):
pubmed: 27852860
J Struct Biol. 2018 Aug;203(2):102-108
pubmed: 29605570
J Biol Chem. 2008 Mar 21;283(12):7877-84
pubmed: 18187419
Structure. 2018 Sep 4;26(9):1187-1195.e4
pubmed: 30017564
Anal Chem. 2012 Mar 20;84(6):2939-48
pubmed: 22409725
Acta Crystallogr D Biol Crystallogr. 2011 Apr;67(Pt 4):235-42
pubmed: 21460441
Protein Sci. 2015 Aug;24(8):1176-92
pubmed: 25676284
Curr HIV Res. 2013 Dec;11(8):623-34
pubmed: 24606219
Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7366-70
pubmed: 1871141
J Biol Chem. 2007 May 25;282(21):15743-53
pubmed: 17403681
Cell. 1991 Apr 19;65(2):241-8
pubmed: 2015625
Cell. 1991 Oct 4;67(1):169-78
pubmed: 1913815
Cell. 2016 Sep 22;167(1):158-170.e12
pubmed: 27662088
Science. 1989 Dec 22;246(4937):1625-9
pubmed: 2688093
Science. 1996 Sep 13;273(5281):1547-51
pubmed: 8703216
Nature. 2011 Dec 21;481(7381):365-70
pubmed: 22190034
PLoS One. 2012;7(11):e48688
pubmed: 23133650
Mol Cell. 2019 Mar 21;73(6):1191-1203.e6
pubmed: 30824373
PLoS Pathog. 2007 Apr;3(4):e54
pubmed: 17432934
J Virol. 1989 Mar;63(3):1265-74
pubmed: 2783738
Cold Spring Harb Perspect Med. 2012 Feb;2(2):a006916
pubmed: 22355797
Methods Enzymol. 2009;455:127-55
pubmed: 19289205
Proc Natl Acad Sci U S A. 1997 Feb 4;94(3):973-8
pubmed: 9023367
Cell. 1989 Jun 30;57(7):1155-65
pubmed: 2736624
Semin Cell Dev Biol. 2018 Mar;75:70-77
pubmed: 28866329
J Virol. 1998 Apr;72(4):2935-44
pubmed: 9525614
Mol Cell Biol. 1999 Feb;19(2):1210-7
pubmed: 9891055
J Mol Biol. 2011 Jul 29;410(5):959-71
pubmed: 21763499
Traffic. 2005 Mar;6(3):187-98
pubmed: 15702987
J Mol Biol. 2018 Feb 16;430(4):537-553
pubmed: 28705764
Retrovirology. 2011 Mar 14;8:17
pubmed: 21401918
Cell. 1999 May 28;97(5):635-46
pubmed: 10367892
Annu Rev Microbiol. 1998;52:491-532
pubmed: 9891806
EMBO J. 1998 Aug 3;17(15):4491-502
pubmed: 9687515
J Virol. 1991 Dec;65(12):7051-5
pubmed: 1942257
Chemphyschem. 2013 Sep 16;14(13):3034-45
pubmed: 23794453
Nat Protoc. 2016 May;11(5):882-94
pubmed: 27055097
Acta Crystallogr D Biol Crystallogr. 2014 Apr;70(Pt 4):1050-60
pubmed: 24699649
Mol Cell. 2008 Sep 26;31(6):824-34
pubmed: 18922466
Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7734-8
pubmed: 1715576
EMBO J. 1996 Apr 15;15(8):1818-25
pubmed: 8617227
Elife. 2014 Aug 27;3:e03656
pubmed: 25163983
J Virol. 2010 Jul;84(13):6598-604
pubmed: 20427541
Curr Biol. 1997 Oct 1;7(10):767-75
pubmed: 9368759
J Mol Biol. 1994 Aug 12;241(2):193-207
pubmed: 8057359
J Cell Biol. 1999 Jan 25;144(2):213-24
pubmed: 9922449
Methods Mol Biol. 2018;1764:133-151
pubmed: 29605913
Nat Struct Mol Biol. 2010 Nov;17(11):1367-76
pubmed: 20972448
Nature. 1986 May 22-28;321(6068):412-7
pubmed: 3012355
Anal Chem. 2006 Nov 1;78(21):7473-83
pubmed: 17073415
Proc Natl Acad Sci U S A. 2010 Mar 30;107(13):5810-4
pubmed: 20231488
J Mol Biol. 2008 Nov 21;383(4):772-82
pubmed: 18708071
J Biol Chem. 2006 Jul 28;281(30):20883-20890
pubmed: 16704975
Wiley Interdiscip Rev RNA. 2016 Jul;7(4):470-86
pubmed: 26929078
J Biomol NMR. 1995 Nov;6(3):277-93
pubmed: 8520220
Semin Cell Dev Biol. 2018 Jul;79:131-142
pubmed: 29102717
Nature. 1999 May 20;399(6733):221-9
pubmed: 10353244
J Struct Biol. 1998 Jan;121(1):41-52
pubmed: 9573619
Cell. 1998 Jul 24;94(2):193-204
pubmed: 9695948
Biochemistry. 1991 Jul 30;30(30):7527-34
pubmed: 1854752
Protein Sci. 2006 Dec;15(12):2795-804
pubmed: 17088319
J Mol Biol. 2005 Jun 10;349(3):515-25
pubmed: 15878174
Genes Dev. 1991 May;5(5):808-19
pubmed: 1827422
J Mol Biol. 2016 Feb 22;428(4):720-725
pubmed: 26410586
Chem Biol. 1999 Sep;6(9):657-69
pubmed: 10467126
Virology. 1994 Jul;202(1):186-94
pubmed: 7516596
Biochim Biophys Acta. 2005 Mar 22;1743(1-2):169-75
pubmed: 15777852
Chembiochem. 2018 Aug 6;19(15):1618-1624
pubmed: 29791766
Proc Natl Acad Sci U S A. 2009 Feb 3;106(5):1404-8
pubmed: 19164515
J Proteomics. 2013 Aug 02;88:120-8
pubmed: 23541715
Nat Struct Biol. 1996 Dec;3(12):1026-33
pubmed: 8946856
Nat Protoc. 2018 Mar;13(3):478-494
pubmed: 29419816
Cell. 1997 Sep 19;90(6):1051-60
pubmed: 9323133
Nucleic Acids Res. 2017 May 5;45(8):4632-4641
pubmed: 28379444
Nature. 1997 Nov 20;390(6657):308-11
pubmed: 9384386
J Biol Chem. 2004 Jan 16;279(3):2176-81
pubmed: 14561738
J Virol. 1990 Nov;64(11):5360-6
pubmed: 2120472
Nat Commun. 2019 Jul 30;10(1):3404
pubmed: 31363125
Nature. 2005 Jun 2;435(7042):693-6
pubmed: 15864302
Genes Dev. 1990 Aug;4(8):1357-64
pubmed: 2227413
Cell. 1986 Sep 12;46(6):807-17
pubmed: 3638988
Structure. 2016 Jul 6;24(7):1068-80
pubmed: 27265851
Biophys J. 2015 Jul 21;109(2):277-86
pubmed: 26200863
Nat Protoc. 2016 Dec;11(12):2301-2319
pubmed: 27809316
J Biol Chem. 2002 Dec 27;277(52):50597-606
pubmed: 12372823
Mol Cell Proteomics. 2016 Jul;15(7):2491-500
pubmed: 27150526
Protein Sci. 2008 Mar;17(3):420-30
pubmed: 18218716
J Mol Biol. 2016 May 22;428(10 Pt A):2060-90
pubmed: 26523678
Mol Cell. 2002 Dec;10(6):1345-53
pubmed: 12504010
Biochemistry. 2003 Mar 11;42(9):2625-33
pubmed: 12614157
J Virol. 1993 Aug;67(8):4769-76
pubmed: 8331728
Biochemistry. 1995 Jul 4;34(26):8242-9
pubmed: 7599117
Structure. 2010 Sep 8;18(9):1171-83
pubmed: 20826343
Biochim Biophys Acta. 2011 Sep;1813(9):1578-92
pubmed: 21029753
J Biol Chem. 2009 Nov 27;284(48):33384-91
pubmed: 19808671
J Mol Biol. 2007 Dec 7;374(4):1129-38
pubmed: 18028944
Science. 2003 Nov 28;302(5650):1571-5
pubmed: 14645851
Sci Rep. 2019 Mar 26;9(1):5139
pubmed: 30914719
Elife. 2014 Dec 08;3:e04120
pubmed: 25486594
Nat Commun. 2015 Jun 10;6:7244
pubmed: 26059226
J Biomol NMR. 2013 Apr;55(4):311-21
pubmed: 23435576
Proc Natl Acad Sci U S A. 2010 Jul 13;107(28):12481-6
pubmed: 20616058
Nucleus. 2010 Mar-Apr;1(2):190-201
pubmed: 21326951
Proc Natl Acad Sci U S A. 1989 Mar;86(5):1495-9
pubmed: 2784208
Anal Chem. 2002 Mar 15;74(6):1402-7
pubmed: 11922310
Structure. 2000 Mar 15;8(3):329-38
pubmed: 10745017
Nat Struct Mol Biol. 2010 Nov;17(11):1337-42
pubmed: 20953181
EMBO J. 1996 Apr 15;15(8):1810-7
pubmed: 8617226
Mol Cell Proteomics. 2012 Apr;11(4):M111.015313
pubmed: 22174317
J Biomol NMR. 2004 Sep;30(1):11-23
pubmed: 15452431
J Mol Biol. 1997 Dec 19;274(5):693-707
pubmed: 9405152
Acta Crystallogr D Struct Biol. 2016 Jun;72(Pt 6):705-17
pubmed: 27303791
Nat Rev Mol Cell Biol. 2007 Mar;8(3):195-208
pubmed: 17287812
Mol Cell Biol. 1992 Mar;12(3):1375-86
pubmed: 1545819
J Virol. 1996 Feb;70(2):1282-7
pubmed: 8551596
Protein Expr Purif. 2005 May;41(1):207-34
pubmed: 15915565
Cell. 2000 Jul 7;102(1):99-108
pubmed: 10929717
Nature. 1989 Mar 16;338(6212):254-7
pubmed: 2784194
Methods. 2015 Apr;76:124-136
pubmed: 25484338