title: NTF2-like domain of Tap plays a critical role in cargo mRNA recognition and export.
abstract: Metazoan Tap-p15 (also called Nxf1-Nxt1) and yeast Mex67-Mtr2 heterodimers are the general mRNA export receptors. The RNA binding activity of Tap-p15, which is essential for mRNA nuclear export, has been attributed to the amino-terminal RNA binding module of Tap consists of RNA recognition motif (RRM) and leucine-rich repeat. In this study, we identified a novel RNA interaction surface in the NTF2-like (NTF2L) domain of Tap, which is analogous to the rRNA binding platform of Mex67-Mtr2. Tap-p15 uses the three domains to tightly bind the retroviral constitutive transport element. The RNA binding through the NTF2L domain is functionally relevant as introduction of mutations in this region reduced CTE-containing mRNA export activity. In contrast, only when the RRM and NTF2L domains were mutated simultaneously, bulk poly (A)(+) RNA export and in vivo poly (A)(+) RNA binding activities of Tap-p15 were significantly attenuated. Moreover, an engineered human cell line harboring the NTF2L domain mutation in the NXF1 gene showed a synthetic growth phenotype and severe mRNA export defect under Aly/REF and Thoc5 depleted condition. These data suggest that Tap-p15 recognizes bulk mRNAs through combinatorial use of the distinct RNA binding domains.
authors: Biomolecular Networks Laboratories, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan Department of Biochemistry, Graduate School of Medicine, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan katahira@anat3.med.osaka-u.ac.jp.
authors: Jun
authors: J
authors: Katahira
authors: Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, Heidelberg D-69120, Germany.
authors: Lyudmila
authors: L
authors: Dimitrova
authors: Department of Biological Informatics and Experimental Therapeutics, Graduate School of Medicine, Akita University, Akita 010-8543, Japan.
authors: Yumiko
authors: Y
authors: Imai
authors: Biochemie-Zentrum der Universität Heidelberg, Im Neuenheimer Feld 328, Heidelberg D-69120, Germany.
authors: Ed
authors: E
authors: Hurt
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D001692:Biological Transport; D002460:Cell Line; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D017931:DNA Primers; D019281:Dimerization; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D029968:Escherichia coli Proteins; D012333:RNA, Messenger; C067124:Tap protein, E coli
references: Genomics. 2005 May;85(5):641-53
references: 15820316
references: Genes Dev. 1999 May 1;13(9):1126-39
references: 10323864
references: EMBO J. 2006 Nov 1;25(21):5126-37
references: 17036044
references: Nat Rev Mol Cell Biol. 2007 Mar;8(3):195-208
references: 17287812
references: Mol Cell. 2007 Apr 13;26(1):51-62
references: 17434126
references: Nat Rev Mol Cell Biol. 2007 Oct;8(10):761-73
references: 17786152
references: Proc Natl Acad Sci U S A. 2008 Apr 1;105(13):5154-9
references: 18364396
references: EMBO J. 2009 Mar 4;28(5):556-67
references: 19165146
references: Nature. 2009 Sep 3;461(7260):60-5
references: 19680239
references: Science. 2009 Nov 27;326(5957):1275-9
references: 19965479
references: Nat Rev Mol Cell Biol. 2010 Jul;11(7):490-501
references: 20571586
references: Cold Spring Harb Perspect Biol. 2010 Oct;2(10):a000562
references: 20630994
references: Trends Biochem Sci. 2010 Nov;35(11):609-17
references: 20719516
references: Curr Opin Struct Biol. 2010 Dec;20(6):782-90
references: 20951026
references: Curr Opin Struct Biol. 2011 Feb;21(1):101-8
references: 21145229
references: Nat Rev Mol Cell Biol. 2011 Jun;12(6):377-84
references: 21602906
references: Nat Struct Mol Biol. 2011 Sep;18(9):990-8
references: 21822283
references: Proc Natl Acad Sci U S A. 2012 Mar 6;109(10):3850-5
references: 22357757
references: Transcription. 2012 Jan-Feb;3(1):8-12
references: 22456314
references: Biochim Biophys Acta. 2012 Jun;1819(6):507-13
references: 22178508
references: Biochim Biophys Acta. 2012 Jun;1819(6):578-92
references: 22406340
references: Nat Commun. 2012;3:1006
references: 22893130
references: Science. 2013 Feb 15;339(6121):823-6
references: 23287722
references: Nat Rev Genet. 2013 Apr;14(4):275-87
references: 23478349
references: Cell. 2013 Mar 28;153(1):112-25
references: 23477864
references: Annu Rev Cell Dev Biol. 1999;15:607-60
references: 10611974
references: EMBO J. 2000 Feb 1;19(3):410-20
references: 10722314
references: RNA. 2000 Apr;6(4):638-50
references: 10786854
references: J Cell Biol. 2000 Aug 21;150(4):695-706
references: 10952996
references: Nature. 2000 Sep 21;407(6802):401-5
references: 11014198
references: EMBO J. 2000 Nov 1;19(21):5587-98
references: 11060011
references: Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):1030-5
references: 11158589
references: Mol Cell Biol. 2001 Jul;21(13):4219-32
references: 11390651
references: Genome Biol. 2001;2(6):REVIEWS3008
references: 11423015
references: Mol Cell. 2001 Sep;8(3):645-56
references: 11583626
references: Mol Cell Biol. 2002 Jan;22(1):245-56
references: 11739738
references: J Biol Chem. 2002 Mar 15;277(11):9242-6
references: 11756420
references: Mol Cell Biol. 2002 Aug;22(15):5405-18
references: 12101235
references: J Cell Biol. 2002 Nov 25;159(4):579-88
references: 12438415
references: RNA. 2003 Jul;9(7):881-91
references: 12810921
references: Nat Struct Mol Biol. 2004 Jun;11(6):558-66
references: 15133499
references: Proc Natl Acad Sci U S A. 2004 Jun 29;101(26):9666-70
references: 15210956
references: EMBO J. 1997 Jun 2;16(11):3256-71
references: 9214641
references: Mol Cell. 1998 Apr;1(5):649-59
references: 9660949
references: Mol Cell Biol. 1998 Nov;18(11):6826-38
references: 9774696
references: EMBO J. 1999 Apr 1;18(7):1953-65
references: 10202158
references: EMBO J. 1999 May 4;18(9):2593-609
references: 10228171
references: Nucleic Acids Res. 2006;34(17):4711-21
references: 16963774