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

Victor C. Hou, Robert Lersch, Sherry L. Gee, Julie L. Ponthier, Annie J. Lo, Michael Wu, Chris W. Turck, Mark Koury, Adrian R. Krainer, Akila Mayeda, John G. Conboy

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

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 (CE16) 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 CE16 can repress splicing of upstream introns, and that mutagenesis or replacement of CE16 can relieve this inhibition. An affinity selection assay with biotinylated CE16 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.

Original languageEnglish
Pages (from-to)6195-6204
Number of pages10
JournalEMBO Journal
Volume21
Issue number22
DOIs
Publication statusPublished - 15-11-2002

Fingerprint

Heterogeneous-Nuclear Ribonucleoprotein Group A-B
Heterogeneous-Nuclear Ribonucleoproteins
Erythropoiesis
RNA Precursors
Exons
Switches
Chemical activation
Transcriptional Silencer Elements
Erythroblasts
Mutagenesis
Erythrocyte Membrane
Introns
Assays
Down-Regulation
RNA
Membranes

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Hou, V. C., Lersch, R., Gee, S. L., Ponthier, J. L., Lo, A. J., Wu, M., ... Conboy, J. G. (2002). Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch. EMBO Journal, 21(22), 6195-6204. https://doi.org/10.1093/emboj/cdf625
Hou, Victor C. ; Lersch, Robert ; Gee, Sherry L. ; Ponthier, Julie L. ; Lo, Annie J. ; Wu, Michael ; Turck, Chris W. ; Koury, Mark ; Krainer, Adrian R. ; Mayeda, Akila ; Conboy, John G. / Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch. In: EMBO Journal. 2002 ; Vol. 21, No. 22. pp. 6195-6204.
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Hou, VC, Lersch, R, Gee, SL, Ponthier, JL, Lo, AJ, Wu, M, Turck, CW, Koury, M, Krainer, AR, Mayeda, A & Conboy, JG 2002, 'Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch', EMBO Journal, vol. 21, no. 22, pp. 6195-6204. https://doi.org/10.1093/emboj/cdf625

Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch. / Hou, Victor C.; Lersch, Robert; Gee, Sherry L.; Ponthier, Julie L.; Lo, Annie J.; Wu, Michael; Turck, Chris W.; Koury, Mark; Krainer, Adrian R.; Mayeda, Akila; Conboy, John G.

In: EMBO Journal, Vol. 21, No. 22, 15.11.2002, p. 6195-6204.

Research output: Contribution to journalArticle

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T1 - Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch

AU - Hou, Victor C.

AU - Lersch, Robert

AU - Gee, Sherry L.

AU - Ponthier, Julie L.

AU - Lo, Annie J.

AU - Wu, Michael

AU - Turck, Chris W.

AU - Koury, Mark

AU - Krainer, Adrian R.

AU - Mayeda, Akila

AU - Conboy, John G.

PY - 2002/11/15

Y1 - 2002/11/15

N2 - 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 (CE16) 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 CE16 can repress splicing of upstream introns, and that mutagenesis or replacement of CE16 can relieve this inhibition. An affinity selection assay with biotinylated CE16 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.

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Hou VC, Lersch R, Gee SL, Ponthier JL, Lo AJ, Wu M et al. Decrease in hnRNP A/B expression during erythropoiesis mediates a pre-mRNA splicing switch. EMBO Journal. 2002 Nov 15;21(22):6195-6204. https://doi.org/10.1093/emboj/cdf625