Regulated Fox-2 isoform expression mediates protein 4.1R splicing during erythroid differentiation.
A regulated splicing event in protein 4.1R pre-mRNA-the inclusion of exon 16-encoding peptides for spectrin-actin binding-occurs in late erythroid differentiation. We defined the functional significance of an intronic splicing enhancer, UGCAUG, and its cognate splicing factor, mFox2A, on exon 16 splicing during differentiation. UGCAUG displays cell-type-specific splicing regulation in a test neutral reporter and has a dose-dependent enhancing effect. Erythroid cells express 2 UGCAUG-binding mFox-2 isoforms, an erythroid differentiation-inducible mFox-2A and a commonly expressed mFox-2F. When overexpressed, both enhanced internal exon splicing in an UGCAUG-dependent manner, with mFox-2A exerting a much stronger effect than mFox-2F. A significant reciprocal increase in mFox-2A and decrease in mFox-2F occurred during erythroid differentiation and correlated with exon 16 inclusion. Furthermore, isoform-specific expression reduction reversed mFox-2A-enhancing activity, but not that of mFox-2F on exon 16 inclusion. Our results suggest that an erythroid differentiation-inducible mFox-2A isoform is a critical regulator of the differentiation-specific exon 16 splicing switch, and that its up-regulation in late erythroid differentiation is vital for exon 16 splicing.
Yang
Guang
G
Department of Medical Oncology, Dana-Farber Cancer Institute, 44 Binney St, Boston, MA 02115, USA.
Huang
Shu-Ching
SC
Wu
Jane Y
JY
Benz
Edward J
EJ
D000818:Animals; D001798:Blood Proteins / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology; D003598:Cytoskeletal Proteins / Q000235:genetics*; D024202:Electrophoretic Mobility Shift Assay; D041905:Erythroid Cells / Q000166:cytology*; D005091:Exons / Q000235:genetics; D006367:HeLa Cells; D051542:Hepatocyte Nuclear Factor 3-beta / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007438:Introns / Q000235:genetics; D007536:Isomerism; D004915:Leukemia, Erythroblastic, Acute; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008840:Microfilament Proteins; D016296:Mutagenesis; D012322:RNA Precursors / Q000378:metabolism; D012326:RNA Splicing / Q000502:physiology*
D001798:Blood Proteins; D003598:Cytoskeletal Proteins; C514956:Epb41 protein, mouse; C496502:FOXA2 protein, human; D008565:Membrane Proteins; D008840:Microfilament Proteins; D012322:RNA Precursors; C028841:erythrocyte membrane band 4.1 protein; D051542:Hepatocyte Nuclear Factor 3-beta
Mol Cell. 1998 Apr;1(5):765-71
9660960
Mol Cell Biol. 1998 Jul;18(7):3900-6
9632774
Mol Cell Biol. 1999 Mar;19(3):1853-63
10022872
J Cell Biol. 1999 Apr 5;145(1):29-43
10189366
Mol Biol Cell. 2005 Jan;16(1):117-27
15525677
Blood. 2005 Mar 1;105(5):2146-53
15522963
BMC Mol Biol. 2005;6:4
15720708
Nucleic Acids Res. 2005;33(7):2078-89
15824060
EMBO J. 2005 Aug 3;24(15):2792-802
16001081
Nat Methods. 2004 Nov;1(2):163-7
16144086
Mol Cell Biol. 2005 Nov;25(22):10005-16
16260614
Mol Cell Biol. 2006 Feb;26(4):1209-22
16449636
J Biol Chem. 2006 May 5;281(18):12468-74
16537540
Mol Cell Biol. 2007 Feb;27(3):830-41
17101796
Trends Biochem Sci. 2000 Aug;25(8):381-8
10916158
RNA. 2000 Sep;6(9):1197-211
10999598
Nucleic Acids Res. 2001 Jun 1;29(11):2338-48
11376152
Biochem Biophys Res Commun. 2001 Mar 30;282(2):499-506
11401487
Blood. 2001 Dec 15;98(13):3809-16
11739190
Mol Endocrinol. 2002 Mar;16(3):459-68
11875103
EMBO J. 2002 Nov 15;21(22):6195-204
12426391
EMBO J. 2003 Feb 17;22(4):905-12
12574126
Neurosci Lett. 2003 Mar 13;339(1):62-6
12618301
Blood. 2003 May 15;101(10):4164-71
12522012
Annu Rev Biochem. 2003;72:291-336
12626338
Blood. 2003 Dec 1;102(12):3938-46
12907435
Mol Cell Biol. 2003 Dec;23(24):9327-37
14645542
Proc Natl Acad Sci U S A. 1988 Dec;85(23):9062-5
3194408
J Cell Biol. 1990 Mar;110(3):617-24
2307701
Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5222-6
1647028
J Clin Invest. 1993 Jan;91(1):329-38
8423229
Trans Assoc Am Physicians. 1992;105:165-71
1308992
J Cell Sci. 1994 Jan;107 ( Pt 1):1-7
8175901
Trends Genet. 1994 Mar;10(3):100-6
8178363
Blood. 1996 May 1;87(9):3934-41
8611723
Blood. 1996 Jun 15;87(12):5324-31
8652848
J Biol Chem. 1996 Jul 26;271(30):17613-6
8663598
Mol Cell Biol. 1997 Nov;17(11):6537-45
9343417
Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12343-7
9356451
Annu Rev Genet. 1998;32:279-305
9928482