The Neuronal Splicing Factor Nova Co-Localizes with Target RNAs in the Dendrite.
Nova proteins are neuron-specific RNA binding proteins targeted by autoantibodies in a disorder manifest by failure of motor inhibition, and they regulate splicing and alternative 3' processing. Nova regulates splicing of RNAs encoding synaptic proteins, including the inhibitory glycine receptor alpha2 subunit (GlyRalpha2), and binds to others, including the GIRK2 channel. We found that Nova harbors functional NES and NLS elements, shuttles between the nucleus and cytoplasm, and that 50% of the protein localizes to the soma-dendritic compartment. Immunofluoresence and EM analysis of spinal cord motor neurons demonstrated that Nova co-localizes beneath synaptic contacts in dendrites with the same RNA, GlyRalpha2, whose splicing it regulates in the nucleus. HITS-CLIP identified intronic and 3' UTR sites where Nova binds to GlyRalpha2 and GIRK2 transcripts in the brain. This led directly to the identification of a 3' UTR localization element that mediates Nova-dependent localization of GIRK2 in primary neurons. These data demonstrate that HITS-CLIP can identify functional RNA localization elements, and they suggest new links between the regulation of nuclear RNA processing and mRNA localization.
Racca
Claudia
C
Biologie Cellulaire de la Synapse Normale et Pathologique, Institut National de la Santé et de la Recherche Médicale, Ecole Normale Supérieure Paris, France.
Gardiol
Alejandra
A
Eom
Taesun
T
Ule
Jernej
J
Triller
Antoine
A
Darnell
Robert B
RB
Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3513-8
19221030
Annu Rev Neurosci. 2001;24:299-325
11283313
Neuroscience. 1991;43(2-3):381-95
1656320
Cell. 2005 Oct 7;123(1):105-18
16213216
Dev Cell. 2003 Jan;4(1):41-51
12530962
Curr Opin Cell Biol. 2008 Apr;20(2):144-9
18378131
Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):13254-9
9789075
J Cell Biol. 1985 Aug;101(2):683-8
2991304
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5977-81
392511
Cell. 1991 Apr 19;65(2):241-8
2015625
Nature. 2004 Feb 19;427(6976):753-7
14973490
Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6350-5
10829067
Neuron. 2000 Feb;25(2):359-71
10719891
Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):14937-42
10611316
Neuroscience. 1998 Jun;84(4):997-1012
9578391
Cell. 2006 Jan 13;124(1):191-205
16413491
J Neurosci. 2006 Jul 5;26(27):7143-6
16822969
Nature. 2008 Nov 27;456(7221):464-9
18978773
Cell. 2009 Feb 20;136(4):719-30
19239891
Cell. 2000 Feb 4;100(3):323-32
10676814
Annu Rev Cell Dev Biol. 2001;17:569-614
11687499
Nature. 1992 Feb 20;355(6362):730-2
1371331
J Neurosci. 1997 Mar 1;17(5):1691-700
9030628
Science. 2003 Nov 14;302(5648):1212-5
14615540
J Cell Biol. 2004 Apr 26;165(2):203-11
15096527
EMBO J. 2007 Aug 22;26(16):3729-36
17660744
EMBO J. 1991 Sep;10(9):2401-9
1651228
Nat Rev Neurosci. 2001 Apr;2(4):240-50
11283747
EMBO J. 2005 Apr 20;24(8):1608-20
15933722
J Neurosci. 1999 Jan 1;19(1):168-79
9870948
Semin Oncol. 2006 Jun;33(3):270-98
16769417
FEBS Lett. 1991 May 20;283(1):73-7
1645300
Results Probl Cell Differ. 2001;34:41-56
11288678
Mol Cell. 2001 Dec;8(6):1351-61
11779509
Nature. 2006 Nov 30;444(7119):580-6
17065982
Nat Struct Mol Biol. 2009 Oct;16(10):1056-62
19734899
Neuron. 2003 Oct 30;40(3):581-94
14642281
Curr Biol. 2002 Jan 22;12(2):R50-2
11818077
Brain Res. 1986 Mar 26;369(1-2):316-20
3008938
Genes Dev. 2000 May 1;14(9):1098-108
10809668
Trends Biochem Sci. 2007 Jun;32(6):279-85
17481902
J Cell Biol. 1983 May;96(5):1337-54
6404910
Mol Cell Biol. 2003 Jul;23(13):4687-700
12808107
J Cell Biol. 1987 Apr;104(4):947-56
3031086
Annu Rev Neurosci. 2001;24:239-62
11283311
Mol Cell. 2007 Nov 9;28(3):359-70
17996701
Neuron. 2008 Oct 23;60(2):201-14
18957214
Nat Rev Genet. 2010 Jan;11(1):75-87
20019688
Proc Natl Acad Sci U S A. 1984 Nov;81(22):7224-7
6095276
Nature. 2008 Nov 27;456(7221):470-6
18978772
J Neurosci. 2008 Nov 12;28(46):11760-7
19005037
Science. 2007 May 25;316(5828):1191-4
17525339
J Neurosci. 2003 Nov 12;23(32):10433-44
14614102
Semin Cell Dev Biol. 2007 Apr;18(2):186-93
17459736
J Neurosci. 1994 Jan;14(1):39-53
8283246
Nature. 1987 Nov 5-11;330(6143):25-6
2823147
Br J Pharmacol. 2004 Sep;143(1):19-26
15302677
Cell. 1986 May 9;45(3):407-15
3698103
Science. 1995 Sep 22;269(5231):1734-7
7569904
Mol Cell Biol. 1997 Jun;17(6):3194-201
9154818
Cell. 2006 Oct 6;127(1):49-58
17018276
Curr Opin Neurobiol. 2006 Feb;16(1):102-10
16418001
J Neurosci. 1996 Feb 1;16(3):1114-22
8558240
Neuron. 1993 Oct;11(4):657-72
8398153
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5982-6
392512
Methods. 2005 Dec;37(4):376-86
16314267
J Cell Biol. 1997 Mar 24;136(6):1263-70
9087442
Cold Spring Harb Symp Quant Biol. 2006;71:321-7
17381312
Genes Dev. 2009 Jan 1;23(1):1-11
19136621
Nat Struct Mol Biol. 2008 Aug;15(8):768-74
18679428
Mol Cell. 2000 Sep;6(3):673-82
11030346
Neuron. 1990 Dec;5(6):867-73
2176511
Curr Opin Cell Biol. 1992 Dec;4(6):975-8
1485968
N Engl J Med. 2003 Oct 16;349(16):1543-54
14561798
J Cell Biol. 2002 Jan 7;156(1):41-51
11781334
Nature. 2004 Apr 29;428(6986):959-63
15118729
Science. 2009 Nov 27;326(5957):1212-6
19965463
Nat Genet. 2005 Aug;37(8):844-52
16041372
Nat Rev Mol Cell Biol. 2005 May;6(5):363-75
15852043
Curr Biol. 2001 Oct 30;11(21):1666-74
11696323
Methods Mol Biol. 2008;488:85-98
18982285