RBFOX2 Promotes Protein 4.1R Exon 16 Selection Via U1 SnRNP Recruitment

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2011 Nov 16

PMID 22083953

Citations 27

Authors

Shu-Ching Huang

Alexander C Ou

Jennie Park

Faye Yu

Brian Yu

Angela Lee

Guang Yang

Anyu Zhou

Edward J Benz Jr

Affiliations

Soon will be listed here.

Abstract

The erythroid differentiation-specific splicing switch of protein 4.1R exon 16, which encodes a spectrin/actin-binding peptide critical for erythrocyte membrane stability, is modulated by the differentiation-induced splicing factor RBFOX2. We have now characterized the mechanism by which RBFOX2 regulates exon 16 splicing through the downstream intronic element UGCAUG. Exon 16 possesses a weak 5' splice site (GAG/GTTTGT), which when strengthened to a consensus sequence (GAG/GTAAGT) leads to near-total exon 16 inclusion. Impaired RBFOX2 binding reduces exon 16 inclusion in the context of the native weak 5' splice site, but not the engineered strong 5' splice site, implying that RBFOX2 achieves its effect by promoting utilization of the weak 5' splice site. We further demonstrate that RBFOX2 increases U1 snRNP recruitment to the weak 5' splice site through direct interaction between its C-terminal domain (CTD) and the zinc finger region of U1C and that the CTD is required for the effect of RBFOX2 on exon 16 splicing. Our data suggest a novel mechanism for exon 16 5' splice site activation in which the binding of RBFOX2 to downstream intronic splicing enhancers stabilizes the pre-mRNA-U1 snRNP complex through interactions with U1C.

Citing Articles

RBFOX2 as a regulatory linchpin in cancer: insights from a comprehensive review of its roles in tumorigenesis.

Liao S, Zhou Z, Jiao Y, Chen S, Bao Y, Cao J Am J Cancer Res. 2024; 14(10):5045-5060.

PMID: 39553227 PMC: 11560822. DOI: 10.62347/BNPO2363.

Evolution and stress response potential of the plant splicing factor U1C.

Jia Z, Wang J, Meng X, Yang X, Tian Y, Wang B Sci Rep. 2024; 14(1):17212.

PMID: 39060315 PMC: 11282270. DOI: 10.1038/s41598-024-68190-4.

Alternative splicing and related RNA binding proteins in human health and disease.

Tao Y, Zhang Q, Wang H, Yang X, Mu H Signal Transduct Target Ther. 2024; 9(1):26.

PMID: 38302461 PMC: 10835012. DOI: 10.1038/s41392-024-01734-2.

Splicing regulation of GFPT1 muscle-specific isoform and its roles in glucose metabolisms and neuromuscular junction.

Farshadyeganeh P, Nazim M, Zhang R, Ohkawara B, Nakajima K, Rahman M iScience. 2023; 26(10):107746.

PMID: 37744035 PMC: 10514471. DOI: 10.1016/j.isci.2023.107746.

LncRNA-Profile-Based Screening of Extracellular Vesicles Released from Brain Endothelial Cells after Oxygen-Glucose Deprivation.

He X, Zou H, Lyu Q, Tang Y, Xiong W, Shang F Brain Sci. 2022; 12(8).

PMID: 36009090 PMC: 9405926. DOI: 10.3390/brainsci12081027.

References

Jurica M, Moore M . Pre-mRNA splicing: awash in a sea of proteins. Mol Cell. 2003; 12(1):5-14. DOI: 10.1016/s1097-2765(03)00270-3. View

Sergeant K, Bourgeois C, Dalgliesh C, Venables J, Stevenin J, Elliott D . Alternative RNA splicing complexes containing the scaffold attachment factor SAFB2. J Cell Sci. 2007; 120(Pt 2):309-19. DOI: 10.1242/jcs.03344. View

Zhou A, Ou A, Cho A, Benz Jr E, Huang S . Novel splicing factor RBM25 modulates Bcl-x pre-mRNA 5' splice site selection. Mol Cell Biol. 2008; 28(19):5924-36. PMC: 2546994. DOI: 10.1128/MCB.00560-08. View

Brudno M, Gelfand M, Spengler S, Zorn M, Dubchak I, Conboy J . Computational analysis of candidate intron regulatory elements for tissue-specific alternative pre-mRNA splicing. Nucleic Acids Res. 2001; 29(11):2338-48. PMC: 55704. DOI: 10.1093/nar/29.11.2338. View

Ohkura N, Takahashi M, Yaguchi H, Nagamura Y, Tsukada T . Coactivator-associated arginine methyltransferase 1, CARM1, affects pre-mRNA splicing in an isoform-specific manner. J Biol Chem. 2005; 280(32):28927-35. DOI: 10.1074/jbc.M502173200. View

Nilsen T . The spliceosome: no assembly required?. Mol Cell. 2002; 9(1):8-9. DOI: 10.1016/s1097-2765(02)00430-6. View

Nelissen R, Heinrichs V, Habets W, Simons F, Luhrmann R, van Venrooij W . Zinc finger-like structure in U1-specific protein C is essential for specific binding to U1 snRNP. Nucleic Acids Res. 1991; 19(3):449-54. PMC: 333632. DOI: 10.1093/nar/19.3.449. View

Zhu H, Hasman R, Young K, Kedersha N, Lou H . U1 snRNP-dependent function of TIAR in the regulation of alternative RNA processing of the human calcitonin/CGRP pre-mRNA. Mol Cell Biol. 2003; 23(17):5959-71. PMC: 180986. DOI: 10.1128/MCB.23.17.5959-5971.2003. View

Heinrichs V, Bach M, Winkelmann G, Luhrmann R . U1-specific protein C needed for efficient complex formation of U1 snRNP with a 5' splice site. Science. 1990; 247(4938):69-72. DOI: 10.1126/science.2136774. View

10.

Takakuwa Y, Tchernia G, Rossi M, Benabadji M, Mohandas N . Restoration of normal membrane stability to unstable protein 4.1-deficient erythrocyte membranes by incorporation of purified protein 4.1. J Clin Invest. 1986; 78(1):80-5. PMC: 329534. DOI: 10.1172/JCI112577. View

11.

Zahler A, Roth M . Distinct functions of SR proteins in recruitment of U1 small nuclear ribonucleoprotein to alternative 5' splice sites. Proc Natl Acad Sci U S A. 1995; 92(7):2642-6. PMC: 42274. DOI: 10.1073/pnas.92.7.2642. View

12.

Horowitz D, Krainer A . Mechanisms for selecting 5' splice sites in mammalian pre-mRNA splicing. Trends Genet. 1994; 10(3):100-6. DOI: 10.1016/0168-9525(94)90233-x. View

13.

Kawamoto S . Neuron-specific alternative splicing of nonmuscle myosin II heavy chain-B pre-mRNA requires a cis-acting intron sequence. J Biol Chem. 1996; 271(30):17613-6. View

14.

Auweter S, Fasan R, Reymond L, Underwood J, Black D, Pitsch S . Molecular basis of RNA recognition by the human alternative splicing factor Fox-1. EMBO J. 2005; 25(1):163-73. PMC: 1356361. DOI: 10.1038/sj.emboj.7600918. View

15.

Zhang C, Zhang Z, Castle J, Sun S, Johnson J, Krainer A . Defining the regulatory network of the tissue-specific splicing factors Fox-1 and Fox-2. Genes Dev. 2008; 22(18):2550-63. PMC: 2546699. DOI: 10.1101/gad.1703108. View

16.

Fukumura K, Kato A, Jin Y, Ideue T, Hirose T, Kataoka N . Tissue-specific splicing regulator Fox-1 induces exon skipping by interfering E complex formation on the downstream intron of human F1gamma gene. Nucleic Acids Res. 2007; 35(16):5303-11. PMC: 2018636. DOI: 10.1093/nar/gkm569. View

17.

Ponthier J, Schluepen C, Chen W, Lersch R, Gee S, Hou V . Fox-2 splicing factor binds to a conserved intron motif to promote inclusion of protein 4.1R alternative exon 16. J Biol Chem. 2006; 281(18):12468-74. DOI: 10.1074/jbc.M511556200. View

18.

Izquierdo J, Majos N, Bonnal S, Martinez C, Castelo R, Guigo R . Regulation of Fas alternative splicing by antagonistic effects of TIA-1 and PTB on exon definition. Mol Cell. 2005; 19(4):475-84. DOI: 10.1016/j.molcel.2005.06.015. View

19.

Chasis J, Coulombel L, Conboy J, McGee S, Andrews K, Kan Y . Differentiation-associated switches in protein 4.1 expression. Synthesis of multiple structural isoforms during normal human erythropoiesis. J Clin Invest. 1993; 91(1):329-38. PMC: 330030. DOI: 10.1172/JCI116189. View

20.

Yang G, Huang S, Wu J, Benz Jr E . Regulated Fox-2 isoform expression mediates protein 4.1R splicing during erythroid differentiation. Blood. 2007; 111(1):392-401. PMC: 2200819. DOI: 10.1182/blood-2007-01-068940. View