Home LiteratureArticle Details
PMID: 8223481 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Functional domains of the human splicing factor ASF/SF2.

The EMBO journal ·Vol. 12 ·No. 12 ·1993-12-00 ·Pages 4727-37

Zuo P, Manley JL

Abstract

The human splicing factor ASF/SF2 displays two predominant activities in in vitro splicing assays: (i) it is an essential factor apparently required for all splices and (ii) it is able to switch utilization of alternative 5' splice sites in a concentration-dependent manner. ASF/SF2 is the prototype of a family of proteins typified by the presence of one or two RNP-type RNA binding domains (RBDs) and a region highly enriched in repeating arginine-serine dipeptides (RS regions). Here we describe a functional analysis of ASF/SF2, which defines several regions essential for one, or both, of its two principal activities, and provides insights into how this type of protein functions in splicing. Two isoforms of the protein, which arise from alternative splicing, are by themselves inactive, but each can block the activity of ASF/SF2, thereby functioning as splicing repressors. Some, but not all, mutations in the RS region prevent ASF/SF2 from functioning as an essential splicing factor. However, the entire RS region can be deleted without reducing splice site switching activity, indicating that it is not absolutely required for interaction with other splicing factors. Experiments with deletion and substitution mutants reveal that the protein contains two related, but highly diverged, RBDs, and that both are essential for activity. Each RBD by itself retains the ability to bind RNA, although optimal binding requires both domains.

MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Cloning, Molecular DNA Escherichia coli Humans Molecular Sequence Data Nuclear Proteins/antagonists & inhibitors,chemistry,metabolism Point Mutation RNA Splicing RNA-Binding Proteins/chemistry,metabolism Sequence Homology, Amino Acid Serine-Arginine Splicing Factors
Chemicals
Nuclear Proteins RNA-Binding Proteins Serine-Arginine Splicing Factors DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Zuo P
Department of Biological Sciences, Columbia University, New York, NY 10027.
Manley J L
References (50)
50 references, click to expand
  1. A role for exon sequences and splice-site proximity in splice-site selection.
    Cell. 1986 Aug 29;46(5):681-90 PMID: 2427200
  2. The Drosophila RNA-binding protein RBP1 is localized to transcriptionally active sites of chromosomes and shows a functional similarity to human splicing factor ASF/SF2.
    Genes Dev. 1992 Dec;6(12B):2569-79 PMID: 1340470
  3. Splicing of SV40 early pre-mRNA to large T and small t mRNAs utilizes different patterns of lariat branch sites.
    Cell. 1987 Jul 17;50(2):227-36 PMID: 3036371
  4. Regulation of sexual differentiation in D. melanogaster via alternative splicing of RNA from the transformer gene.
    Cell. 1987 Aug 28;50(5):739-47 PMID: 2441872
  5. A factor, U2AF, is required for U2 snRNP binding and splicing complex assembly.
    Cell. 1988 Jan 29;52(2):207-19 PMID: 2963698
  6. Developmental expression of a regulatory gene is programmed at the level of splicing.
    EMBO J. 1987 Dec 20;6(13):4095-104 PMID: 2832151
  7. The sex-determining gene tra-2 of Drosophila encodes a putative RNA binding protein.
    Cell. 1988 Dec 23;55(6):1025-35 PMID: 3144434
  8. Sex-lethal, a Drosophila sex determination switch gene, exhibits sex-specific RNA splicing and sequence similarity to RNA binding proteins.
    Cell. 1988 Dec 23;55(6):1037-46 PMID: 3144435
  9. Alternative splicing of SV40 early pre-mRNA is determined by branch site selection.
    Genes Dev. 1988 Nov;2(11):1460-75 PMID: 2850262
  10. The sex determination locus transformer-2 of Drosophila encodes a polypeptide with similarity to RNA binding proteins.
    Cell. 1989 Mar 24;56(6):1011-8 PMID: 2493992
  11. A common RNA recognition motif identified within a defined U1 RNA binding domain of the 70K U1 snRNP protein.
    Cell. 1989 Apr 7;57(1):89-101 PMID: 2467746
  12. RNA-binding proteins as developmental regulators.
    Genes Dev. 1989 Apr;3(4):431-7 PMID: 2470643
  13. Identification, purification, and biochemical characterization of U2 small nuclear ribonucleoprotein auxiliary factor.
    Proc Natl Acad Sci U S A. 1989 Dec;86(23):9243-7 PMID: 2531895
  14. Structure and expression of the Drosophila melanogaster gene for the U1 small nuclear ribonucleoprotein particle 70K protein.
    Mol Cell Biol. 1990 Jun;10(6):2492-502 PMID: 1692955
  15. A protein factor, ASF, controls cell-specific alternative splicing of SV40 early pre-mRNA in vitro.
    Cell. 1990 Jul 13;62(1):25-34 PMID: 2163768
  16. The essential pre-mRNA splicing factor SF2 influences 5' splice site selection by activating proximal sites.
    Cell. 1990 Jul 13;62(1):35-42 PMID: 2364434
  17. The U2B'' RNP motif as a site of protein-protein interaction.
    EMBO J. 1990 Nov;9(11):3675-81 PMID: 2145152
  18. The Xenopus laevis poly(A) binding protein is composed of multiple functionally independent RNA binding domains.
    EMBO J. 1990 Nov;9(11):3699-705 PMID: 2209558
  19. Purification and characterization of pre-mRNA splicing factor SF2 from HeLa cells.
    Genes Dev. 1990 Jul;4(7):1158-71 PMID: 2145194
  20. Crystal structure of the RNA-binding domain of the U1 small nuclear ribonucleoprotein A.
    Nature. 1990 Dec 6;348(6301):515-20 PMID: 2147232
  21. Recognition of U1 and U2 small nuclear RNAs can be altered by a 5-amino-acid segment in the U2 small nuclear ribonucleoprotein particle (snRNP) B" protein and through interactions with U2 snRNP-A' protein.
    Mol Cell Biol. 1991 Apr;11(4):1829-39 PMID: 1826042
  22. RNA-binding domain of the A protein component of the U1 small nuclear ribonucleoprotein analyzed by NMR spectroscopy is structurally similar to ribosomal proteins.
    Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2495-9 PMID: 1826055
  23. Pre-mRNA splicing in yeast.
    Trends Genet. 1991 Mar;7(3):79-85 PMID: 2031287
  24. The multiple RNA-binding domains of the mRNA poly(A)-binding protein have different RNA-binding activities.
    Mol Cell Biol. 1991 Jul;11(7):3419-24 PMID: 1675426
  25. Characterization of cDNAs encoding the polypyrimidine tract-binding protein.
    Genes Dev. 1991 Jul;5(7):1224-36 PMID: 1906035
  26. Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein.
    Science. 1991 Jul 12;253(5016):157-63 PMID: 1853200
  27. Primary structure of the human splicing factor ASF reveals similarities with Drosophila regulators.
    Cell. 1991 Jul 26;66(2):373-82 PMID: 1855257
  28. Functional expression of cloned human splicing factor SF2: homology to RNA-binding proteins, U1 70K, and Drosophila splicing regulators.
    Cell. 1991 Jul 26;66(2):383-94 PMID: 1830244
  29. RNA recognition: towards identifying determinants of specificity.
    Trends Biochem Sci. 1991 Jun;16(6):214-20 PMID: 1716386
  30. Arginine/serine-rich domains of the su(wa) and tra RNA processing regulators target proteins to a subnuclear compartment implicated in splicing.
    Cell. 1991 Oct 18;67(2):335-42 PMID: 1655279
  31. A novel protein factor is required for use of distal alternative 5' splice sites in vitro.
    Mol Cell Biol. 1991 Dec;11(12):5945-53 PMID: 1658620
  32. Regulation of alternative pre-mRNA splicing by hnRNP A1 and splicing factor SF2.
    Cell. 1992 Jan 24;68(2):365-75 PMID: 1531115
  33. Two members of a conserved family of nuclear phosphoproteins are involved in pre-mRNA splicing.
    Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1301-4 PMID: 1741384
  34. Cloning and domain structure of the mammalian splicing factor U2AF.
    Nature. 1992 Feb 13;355(6361):609-14 PMID: 1538748
  35. The 35-kDa mammalian splicing factor SC35 mediates specific interactions between U1 and U2 small nuclear ribonucleoprotein particles at the 3' splice site.
    Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1725-9 PMID: 1531875
  36. Biochemical mechanisms of constitutive and regulated pre-mRNA splicing.
    Annu Rev Cell Biol. 1991;7:559-99 PMID: 1839712
  37. Isolation of a complementary DNA that encodes the mammalian splicing factor SC35.
    Science. 1992 Apr 24;256(5056):535-8 PMID: 1373910
  38. SR proteins: a conserved family of pre-mRNA splicing factors.
    Genes Dev. 1992 May;6(5):837-47 PMID: 1577277
  39. Multiple activities of the human splicing factor ASF.
    Gene Expr. 1992;2(1):19-29 PMID: 1535526
  40. 1H, 13C, and 15N NMR assignments and global folding pattern of the RNA-binding domain of the human hnRNP C proteins.
    Biochemistry. 1992 Jul 14;31(27):6254-65 PMID: 1385725
  41. Binding of the Drosophila transformer and transformer-2 proteins to the regulatory elements of doublesex primary transcript for sex-specific RNA processing.
    Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8092-6 PMID: 1518835
  42. Multiple factors including the small nuclear ribonucleoproteins U1 and U2 are necessary for pre-mRNA splicing in vitro.
    Cell. 1985 Oct;42(3):725-36 PMID: 2996774
  43. Heterogeneous nuclear ribonucleoproteins: role in RNA splicing.
    Science. 1986 Mar 28;231(4745):1534-9 PMID: 3952495
  44. A single gene from yeast for both nuclear and cytoplasmic polyadenylate-binding proteins: domain structure and expression.
    Cell. 1986 Jun 20;45(6):827-35 PMID: 3518950
  45. General splicing factors SF2 and SC35 have equivalent activities in vitro, and both affect alternative 5' and 3' splice site selection.
    Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11224-8 PMID: 1454802
  46. Cloning and characterization of PSF, a novel pre-mRNA splicing factor.
    Genes Dev. 1993 Mar;7(3):393-406 PMID: 8449401
  47. Distinct functions of SR proteins in alternative pre-mRNA splicing.
    Science. 1993 Apr 9;260(5105):219-22 PMID: 8385799
  48. Identification of an snRNP-associated kinase activity that phosphorylates arginine/serine rich domains typical of splicing factors.
    Nucleic Acids Res. 1993 Jun 25;21(12):2815-22 PMID: 8332490
  49. The basic region of AP-1 specifies glucocorticoid receptor activity at a composite response element.
    Genes Dev. 1992 Dec;6(12B):2491-501 PMID: 1340464
  50. Factors influencing alternative splice site utilization in vivo.
    Mol Cell Biol. 1987 Feb;7(2):738-48 PMID: 3029566
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-12-00
Pages
4727-37
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413918
Subset
IM
Grants
NIGMS NIH HHS · GM 48259 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2