Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch

被引:57
作者
Hou, VC
Lersch, R
Gee, SL
Ponthier, JL
Lo, AJ
Wu, M
Turck, CW
Koury, M
Krainer, AR
Mayeda, A
Conboy, JG [1 ]
机构
[1] Univ Calif Berkeley, Lawrence Berkeley Lab, Div Life Sci, Berkeley, CA 94720 USA
[2] Univ Calif Berkeley, Dept Mol & Cellular Biol, Berkeley, CA 94720 USA
[3] Univ Calif San Francisco, Howard Hughes Med Inst, Dept Med, San Francisco, CA 94143 USA
[4] Univ Calif San Francisco, Cardiovasc Res Inst, San Francisco, CA 94143 USA
[5] Vanderbilt Univ, Dept Med, Vet Affairs Med Ctr, Nashville, TN 37232 USA
[6] Cold Spring Harbor Lab, Cold Spring Harbor, NY 11724 USA
[7] Univ Miami, Sch Med, Dept Biochem & Mol Biol, Miami, FL 33136 USA
关键词
alternative splicing; exonic splicing silencer; hnRNP A and B; protein; 4.1R;
D O I
10.1093/emboj/cdf625
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A physiologically important alternative pre-mRNA splicing switch, involving activation. of protein 4.1R exon 16 (E16) splicing; is required for the establishment of proper mechanical integrity of the erythrocyte membrane during erythropoiesis. Here we identify a conserved exonic splicing silencer element (CE(16)) in E16 that interacts with hnRNP A/B proteins and plays a role in repression of E16 splicing during early erythropoiesis. Experiments with model pre-mRNAs showed that CE(16) can repress splicing of upstream introns, and that mutagenesis or replacement of CE(16) can relieve this inhibition. An affinity selection assay with biotinylated CE(16) RNA demonstrated specific binding of hnRNP A/B proteins. Depletion of hnRNP A/B proteins from nuclear extract significantly increased E16 inclusion, while repletion with recombinant hnRNP A/B restored E16 silencing. Most importantly, differentiating mouse erythroblasts exhibited a stage-specific activation of the E16 splicing switch in concert with a dramatic and specific down-regulation of hnRNP A/B protein expression. These findings demonstrate that natural developmental changes in hnRNP A/B proteins can effect physiologically important switches in pre-mRNA splicing.
引用
收藏
页码:6195 / 6204
页数:10
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