A'-form RNA helices are required for cytoplasmic mRNA transport in Drosophila.
Microtubule-based mRNA transport is widely used to restrict protein expression to specific regions in the cell and has important roles in defining cell polarity and axis determination as well as in neuronal function. However, the structural basis of recognition of cis-acting mRNA localization signals by motor complexes is poorly understood. We have used NMR spectroscopy to describe the first tertiary structure to our knowledge of an RNA element responsible for mRNA transport. The Drosophila melanogaster fs(1)K10 signal, which mediates transport by the dynein motor, forms a stem loop with two double-stranded RNA helices adopting an unusual A'-form conformation with widened major grooves reminiscent of those in B-form DNA. Structure determination of four mutant RNAs and extensive functional assays in Drosophila embryos indicate that the two spatially registered A'-form helices represent critical recognition sites for the transport machinery. Our study provides insights into the basis for RNA cargo recognition and reveals a key biological function encoded by A'-form RNA conformation.
Bullock
Simon L
SL
Division of Cell Biology, Medical Research Council Laboratory of Molecular Biology, Cambridge, UK.
Ringel
Inbal
I
Ish-Horowicz
David
D
Lukavsky
Peter J
PJ
D000818:Animals; D001483:Base Sequence; D003593:Cytoplasm / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000235:genetics; D009690:Nucleic Acid Conformation; D034443:RNA Transport; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors
D029721:Drosophila Proteins; D020409:Molecular Motor Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; C088225:fs(1)K10 protein, Drosophila; D004398:Dyneins
Cell. 1997 Nov 28;91(5):705-12
9393863
Nature. 1990 Aug 16;346(6285):680-2
1696688
Curr Biol. 2006 Jul 25;16(14):1447-52
16860745
J Mol Graph. 1996 Feb;14(1):51-5, 29-32
8744573
Nucleic Acids Res. 2009 Apr;37(6):1713-25
19190094
Nucleic Acids Res. 1999 Feb 15;27(4):949-55
9927725
J Mol Biol. 1997 Mar 28;267(2):338-51
9096230
Methods Enzymol. 2000;317:18-38
10829270
Nat Struct Biol. 1999 Nov;6(11):1055-61
10542099
Science. 1997 Nov 7;278(5340):1111-4
9353189
J Biomol Struct Dyn. 1989 Feb;6(4):655-67
2619933
Nature. 2001 Dec 6;414(6864):611-6
11740552
J Biomol NMR. 2001 Aug;20(4):311-23
11563555
Science. 2009 Nov 27;326(5957):1212-6
19965463
RNA. 2010 Mar;16(3):647-53
20100812
Cell. 2001 Apr 20;105(2):209-19
11336671
Methods Enzymol. 2005;394:399-416
15808230
Development. 1995 Nov;121(11):3809-18
8582290
Mol Cell Biol. 1998 Jul;18(7):3788-95
9632762
Science. 2006 Sep 29;313(5795):1935-42
16959973
Development. 1992 Mar;114(3):653-61
1377623
Cell. 2009 Feb 20;136(4):719-30
19239891
J Neurosci. 1993 Oct;13(10):4214-9
7692010
RNA. 2005 Jul;11(7):1017-29
15987813
Dev Cell. 2005 Jul;9(1):51-62
15992540
Proc Natl Acad Sci U S A. 1976 Mar;73(3):804-8
1062791
Mol Cell Biol. 2005 Feb;25(4):1501-10
15684399
Development. 2008 Jan;135(1):133-43
18045835
RNA. 2004 May;10(5):889-93
15100443
Nat Struct Biol. 2003 Dec;10(12):1033-8
14578934
Biochem Biophys Res Commun. 1972 Sep 26;48(6):1392-9
5077825
Methods Enzymol. 2001;338:320-41
11460556
Genes Dev. 2009 Jul 1;23(13):1546-58
19515976
Methods. 2001 Nov;25(3):316-32
11860286
J Magn Reson. 1999 Jun;138(2):334-42
10341140
Methods Enzymol. 2000;317:220-40
10829283
EMBO J. 2003 May 15;22(10):2484-94
12743042
J Magn Reson. 2003 Jan;160(1):65-73
12565051