Nuclear localization of cytoplasmic poly(A)-binding protein upon rotavirus infection involves the interaction of NSP3 with eIF4G and RoXaN.
Rotavirus nonstructural protein NSP3 interacts specifically with the 3' end of viral mRNAs, with the eukaryotic translation initiation factor eIF4G, and with RoXaN, a cellular protein of yet-unknown function. By evicting cytoplasmic poly(A) binding protein (PABP-C1) from translation initiation complexes, NSP3 shuts off the translation of cellular polyadenylated mRNAs. We show here that PABP-C1 evicted from eIF4G by NSP3 accumulates in the nucleus of rotavirus-infected cells. Through modeling of the NSP3-RoXaN complex, we have identified mutations in NSP3 predicted to interrupt its interaction with RoXaN without disturbing the NSP3 interaction with eIF4G. Using these NSP3 mutants and a deletion mutant unable to associate with eIF4G, we show that the nuclear localization of PABP-C1 not only is dependent on the capacity of NSP3 to interact with eIF4G but also requires the interaction of NSP3 with a specific region in RoXaN, the leucine- and aspartic acid-rich (LD) domain. Furthermore, we show that the RoXaN LD domain functions as a nuclear export signal and that RoXaN tethers PABP-C1 with RNA. This work identifies RoXaN as a cellular partner of NSP3 involved in the nucleocytoplasmic localization of PABP-C1.
Harb
Maya
M
Virologie Moléculaire et Structurale INRA UMR 1157, CNRS UMR 2472, IFR 115, Avenue de la Terrasse, 91198 Gif sur Yvette, France.
Becker
Michelle M
MM
Vitour
Damien
D
Baron
Carolina H
CH
Vende
Patrice
P
Brown
Spencer C
SC
Bolte
Susanne
S
Arold
Stefan T
ST
Poncet
Didier
D
D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D039603:Eukaryotic Initiation Factor-4G / Q000378:metabolism*; D000882:Haplorhini; D006801:Humans; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D039102:Poly(A)-Binding Proteins / Q000378:metabolism*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D021381:Protein Transport; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012401:Rotavirus / Q000502:physiology*; D017384:Sequence Deletion; D017361:Viral Nonstructural Proteins / Q000378:metabolism*
D039603:Eukaryotic Initiation Factor-4G; D050505:Mutant Proteins; C082740:NSP3 protein, Rotavirus; D039102:Poly(A)-Binding Proteins; D016601:RNA-Binding Proteins; D017361:Viral Nonstructural Proteins; C484647:ZC3H7B protein, human
Nucleic Acids Res. 2007;35(18):6017-28
17766253
Cell Death Differ. 2000 Jul;7(7):628-36
10889507
J Virol. 1993 Jun;67(6):3159-65
8388495
J Biol Chem. 2002 Dec 27;277(52):50286-92
12381739
J Virol. 2008 Feb;82(3):1496-504
18032499
Bioinformatics. 2004 Dec 12;20(18):3694-6
15271783
Biochim Biophys Acta. 2004 May 25;1678(2-3):67-84
15157733
Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7029-36
11416183
Mol Cell Biol. 2006 Apr;26(8):3085-97
16581783
J Cell Biol. 2000 Dec 11;151(6):1257-68
11121440
J Biol Chem. 2001 Nov 16;276(46):43083-6
11571283
J Cell Biol. 1995 May;129(3):551-60
7730395
Structure. 2003 Oct;11(10):1207-17
14527389
J Cell Sci. 2000 Dec;113 Pt 23:4139-40
11069756
Virology. 2002 Jun 20;298(1):1-7
12093167
Nat Rev Mol Cell Biol. 2007 Oct;8(10):761-73
17786152
Acta Crystallogr D Biol Crystallogr. 1997 May 1;53(Pt 3):240-55
15299926
RNA. 2003 Dec;9(12):1476-90
14624004
Nucleic Acids Res. 2000 Nov 1;28(21):4068-75
11058101
J Virol. 2004 Apr;78(8):3851-62
15047801
Nat Rev Microbiol. 2007 Jul;5(7):529-39
17571094
Genome Biol. 2003;4(7):223
12844354
J Biol Chem. 2007 Aug 24;282(34):25067-75
17595167
Biochem Soc Trans. 2002 Nov;30(Pt 6):963-9
12440955
Virology. 2005 Apr 10;334(2):284-93
15780878
FEBS Lett. 2002 Feb 20;513(1):114-8
11911889
Mol Cell. 2002 Jun;9(6):1273-83
12086624
Biol Cell. 2002 Sep;94(4-5):205-16
12489690
Methods Enzymol. 2003;374:461-91
14696385
J Virol. 1998 Dec;72(12):9865-72
9811722
J Biol Chem. 1998 May 22;273(21):13015-21
9582337
Structure. 2002 Mar;10(3):319-27
12005431
Genes Dev. 2005 Jan 1;19(1):104-13
15630022
Mol Cell. 2001 Jan;7(1):205-16
11172725
Mol Cell Biol. 2005 May;25(9):3763-73
15831480
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
15572765
Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):981-5
12552102
J Virol. 1997 Jan;71(1):34-41
8985320
J Biol Chem. 2002 Feb 22;277(8):6428-37
11704675
Proc Natl Acad Sci U S A. 2006 Mar 21;103(12):4646-51
16537420
Nucleic Acids Res. 2005 Dec 14;33(22):7074-89
16356927
J Virol. 2000 Aug;74(15):7064-71
10888646
EMBO J. 1998 Oct 1;17(19):5811-21
9755181
Cell Host Microbe. 2007 Apr 19;1(2):147-57
18005692
J Virol. 1996 Feb;70(2):985-91
8551639
J Biol Chem. 2000 Oct 13;275(41):32268-76
10922367
EMBO J. 2006 May 3;25(9):1934-44
16601676
EMBO J. 1998 Dec 15;17(24):7480-9
9857202
Exp Cell Res. 1994 Apr;211(2):400-7
7908267
Trends Cell Biol. 2007 Apr;17(4):193-201
17317185
Nature. 1998 Apr 2;392(6675):520-3
9548260
Emerg Infect Dis. 2006 Feb;12(2):304-6
16494759
J Virol. 1999 Jul;73(7):5411-21
10364288