Sam68 RNA Binding Protein is an in Vivo Substrate for Protein Arginine N-methyltransferase 1

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2003 Jan 17

PMID 12529443

Citations 103

Authors

Jocelyn Cote

Francois-Michel Boisvert

Marie-Chloe Boulanger

Mark T Bedford

Stephane Richard

Affiliations

Soon will be listed here.

Abstract

RNA binding proteins often contain multiple arginine glycine repeats, a sequence that is frequently methylated by protein arginine methyltransferases. The role of this posttranslational modification in the life cycle of RNA binding proteins is not well understood. Herein, we report that Sam68, a heteronuclear ribonucleoprotein K homology domain containing RNA binding protein, associates with and is methylated in vivo by the protein arginine N-methyltransferase 1 (PRMT1). Sam68 contains asymmetrical dimethylarginines near its proline motif P3 as assessed by using a novel asymmetrical dimethylarginine-specific antibody and mass spectrometry. Deletion of the methylation sites and the use of methylase inhibitors resulted in Sam68 accumulation in the cytoplasm. Sam68 was also detected in the cytoplasm of PRMT1-deficient embryonic stem cells. Although the cellular function of Sam68 is unknown, it has been shown to export unspliced human immunodeficiency virus RNAs. Cells treated with methylase inhibitors prevented the ability of Sam68 to export unspliced human immunodeficiency virus RNAs. Other K homology domain RNA binding proteins, including SLM-1, SLM-2, QKI-5, GRP33, and heteronuclear ribonucleoprotein K were also methylated in vivo. These findings demonstrate that RNA binding proteins are in vivo substrates for PRMT1, and their methylation is essential for their proper localization and function.

Citing Articles

Assessment of PRMT6-dependent alternative splicing in pluripotent and differentiating NT2/D1 cells.

Eudenbach M, Busam J, Bouchard C, Rossbach O, Zarnack K, Bauer U Life Sci Alliance. 2025; 8(4).

PMID: 39900436 PMC: 11791029. DOI: 10.26508/lsa.202402946.

Role of Sam68 as an adaptor protein in inflammatory signaling.

Gowd V, Kass J, Sarkar N, Ramakrishnan P Cell Mol Life Sci. 2024; 81(1):89.

PMID: 38351330 PMC: 10864426. DOI: 10.1007/s00018-023-05108-9.

Protein arginine methylation in viral infection and antiviral immunity.

Zheng K, Chen S, Ren Z, Wang Y Int J Biol Sci. 2023; 19(16):5292-5318.

PMID: 37928266 PMC: 10620831. DOI: 10.7150/ijbs.89498.

Arginine methylation of RNA-binding proteins is impaired in Huntington's disease.

Ratovitski T, Kamath S, OMeally R, Gosala K, Holland C, Jiang M Hum Mol Genet. 2023; 32(20):3006-3025.

PMID: 37535888 PMC: 10549789. DOI: 10.1093/hmg/ddad125.

Asymmetric Dimethylation of Ribosomal S6 Kinase 2 Regulates Its Cellular Localisation and Pro-Survival Function.

Khalil M, Ismail H, Panasyuk G, Bdzhola A, Filonenko V, Gout I Int J Mol Sci. 2023; 24(10).

PMID: 37240151 PMC: 10218830. DOI: 10.3390/ijms24108806.

References

McBride A, Weiss V, Kim H, Hogle J, Silver P . Analysis of the yeast arginine methyltransferase Hmt1p/Rmt1p and its in vivo function. Cofactor binding and substrate interactions. J Biol Chem. 2000; 275(5):3128-36. DOI: 10.1074/jbc.275.5.3128. View

Di Fruscio M, Chen T, Richard S . Characterization of Sam68-like mammalian proteins SLM-1 and SLM-2: SLM-1 is a Src substrate during mitosis. Proc Natl Acad Sci U S A. 1999; 96(6):2710-5. PMC: 15834. DOI: 10.1073/pnas.96.6.2710. View

Bedford M, Frankel A, Yaffe M, Clarke S, Leder P, Richard S . Arginine methylation inhibits the binding of proline-rich ligands to Src homology 3, but not WW, domains. J Biol Chem. 2000; 275(21):16030-6. DOI: 10.1074/jbc.M909368199. View

Brahms H, Raymackers J, Union A, de Keyser F, Meheus L, Luhrmann R . The C-terminal RG dipeptide repeats of the spliceosomal Sm proteins D1 and D3 contain symmetrical dimethylarginines, which form a major B-cell epitope for anti-Sm autoantibodies. J Biol Chem. 2000; 275(22):17122-9. DOI: 10.1074/jbc.M000300200. View

Pawlak M, Scherer C, Chen J, Roshon M, Ruley H . Arginine N-methyltransferase 1 is required for early postimplantation mouse development, but cells deficient in the enzyme are viable. Mol Cell Biol. 2000; 20(13):4859-69. PMC: 85937. DOI: 10.1128/MCB.20.13.4859-4869.2000. View

Tang J, Kao P, Herschman H . Protein-arginine methyltransferase I, the predominant protein-arginine methyltransferase in cells, interacts with and is regulated by interleukin enhancer-binding factor 3. J Biol Chem. 2000; 275(26):19866-76. DOI: 10.1074/jbc.M000023200. View

Yun C, Fu X . Conserved SR protein kinase functions in nuclear import and its action is counteracted by arginine methylation in Saccharomyces cerevisiae. J Cell Biol. 2000; 150(4):707-18. PMC: 2175287. DOI: 10.1083/jcb.150.4.707. View

Mowen K, Tang J, Zhu W, Schurter B, Shuai K, Herschman H . Arginine methylation of STAT1 modulates IFNalpha/beta-induced transcription. Cell. 2001; 104(5):731-41. DOI: 10.1016/s0092-8674(01)00269-0. View

Schurter B, Koh S, Chen D, Bunick G, Harp J, Hanson B . Methylation of histone H3 by coactivator-associated arginine methyltransferase 1. Biochemistry. 2001; 40(19):5747-56. DOI: 10.1021/bi002631b. View

10.

FRIESEN W, Massenet S, Paushkin S, Wyce A, Dreyfuss G . SMN, the product of the spinal muscular atrophy gene, binds preferentially to dimethylarginine-containing protein targets. Mol Cell. 2001; 7(5):1111-7. DOI: 10.1016/s1097-2765(01)00244-1. View

11.

Strahl B, Briggs S, Brame C, Caldwell J, Koh S, Ma H . Methylation of histone H4 at arginine 3 occurs in vivo and is mediated by the nuclear receptor coactivator PRMT1. Curr Biol. 2001; 11(12):996-1000. DOI: 10.1016/s0960-9822(01)00294-9. View

12.

McBride A, Silver P . State of the arg: protein methylation at arginine comes of age. Cell. 2001; 106(1):5-8. DOI: 10.1016/s0092-8674(01)00423-8. View

13.

Wang H, Huang Z, Xia L, Feng Q, Erdjument-Bromage H, Strahl B . Methylation of histone H4 at arginine 3 facilitating transcriptional activation by nuclear hormone receptor. Science. 2001; 293(5531):853-7. DOI: 10.1126/science.1060781. View

14.

Chen T, Cote J, Carvajal H, Richard S . Identification of Sam68 arginine glycine-rich sequences capable of conferring nonspecific RNA binding to the GSG domain. J Biol Chem. 2001; 276(33):30803-11. DOI: 10.1074/jbc.M102247200. View

15.

Raman B, Guarnaccia C, Nadassy K, Zakhariev S, Pintar A, Zanuttin F . N(omega)-arginine dimethylation modulates the interaction between a Gly/Arg-rich peptide from human nucleolin and nucleic acids. Nucleic Acids Res. 2001; 29(16):3377-84. PMC: 55848. DOI: 10.1093/nar/29.16.3377. View

16.

FRIESEN W, Paushkin S, Wyce A, Massenet S, Pesiridis G, Van Duyne G . The methylosome, a 20S complex containing JBP1 and pICln, produces dimethylarginine-modified Sm proteins. Mol Cell Biol. 2001; 21(24):8289-300. PMC: 99994. DOI: 10.1128/MCB.21.24.8289-8300.2001. View

17.

Darnell J, Jensen K, Jin P, Brown V, Warren S, Darnell R . Fragile X mental retardation protein targets G quartet mRNAs important for neuronal function. Cell. 2001; 107(4):489-99. DOI: 10.1016/s0092-8674(01)00566-9. View

18.

Brahms H, Meheus L, de Brabandere V, Fischer U, Luhrmann R . Symmetrical dimethylation of arginine residues in spliceosomal Sm protein B/B' and the Sm-like protein LSm4, and their interaction with the SMN protein. RNA. 2001; 7(11):1531-42. PMC: 1370196. DOI: 10.1017/s135583820101442x. View

19.

Xu W, Chen H, Du K, Asahara H, Tini M, Emerson B . A transcriptional switch mediated by cofactor methylation. Science. 2001; 294(5551):2507-11. DOI: 10.1126/science.1065961. View

20.

Frankel A, Yadav N, Lee J, Branscombe T, Clarke S, Bedford M . The novel human protein arginine N-methyltransferase PRMT6 is a nuclear enzyme displaying unique substrate specificity. J Biol Chem. 2001; 277(5):3537-43. DOI: 10.1074/jbc.M108786200. View