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PMID: 8551577 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

SR protein splicing factors interact with the Rous sarcoma virus negative regulator of splicing element.

Journal of virology ·Vol. 70 ·No. 2 ·1996-02-00 ·Pages 1163-72

McNally LM, McNally MT

Abstract

Retroviral replication requires that a portion of the primary transcripts generated from proviral DNA be spliced to serve as mRNA for the envelope protein and in Rous sarcoma virus as src mRNA. However, a substantial amount of full-length RNA must be maintained in an unspliced form, as the unspliced RNA serves both as mRNA for structural proteins and virion-associated enzymatic proteins and as genomic RNA for progeny virions. The extent of viral RNA splicing must be finely controlled, since only a narrow range in the ratio of unspliced RNA to spliced RNA is tolerated for optimal replication. A number of cis-acting sequences within the RNA of Rous sarcoma virus play a role in preserving a large pool of unspliced RNA. One such sequence, the negative regulator of splicing (NRS), is of interest because it blocks splicing but is not located near any of the splice junctions. To better understand how this novel element blocks splicing at a distance, we set out to identify host cell factors that interact specifically with this inhibitory sequence. In this study, proteins from nuclear extracts with molecular masses of 26, 36, 44, and 55 kDa were shown by UV cross-linking assays to bind the NRS preferentially. One of them, p55, was also detected in a specific complex with SR protein electrophoretic mobility shift assay. All but p55 have biochemical properties consistent with SR protein splicing factors, and some, but not all, of the total SR proteins purified from HeLa cells cross-link specifically to the NRS. The strongest cross-linking SR protein is SRp30a/b, which is composed of the splicing factors SF2/ASF and SC35. The NRS specifically binds bacterially expressed SF2/ASF, whereas nonfunctional mutants do not. Data indicating that the 36-kDa protein which cross-links in nuclear extracts is SF2/ASF are presented. The data indicate that factors normally required for RNA splicing may be exploited by retroviruses to block splicing.

MeSH Terms
Avian Sarcoma Viruses/genetics Base Sequence Binding Sites Cross-Linking Reagents HeLa Cells Humans Molecular Sequence Data Nuclear Proteins/metabolism Phosphoproteins/metabolism Purines/metabolism RNA Splicing RNA, Viral/metabolism RNA-Binding Proteins Serine-Arginine Splicing Factors Ultraviolet Rays
Chemicals
Cross-Linking Reagents Nuclear Proteins Phosphoproteins Purines RNA, Viral RNA-Binding Proteins Serine-Arginine Splicing Factors purine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
McNally L M
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226, USA.
McNally M T
References (45)
45 references, click to expand
  1. Identification and purification of a 62,000-dalton protein that binds specifically to the polypyrimidine tract of introns.
    Genes Dev. 1989 Dec;3(12A):1874-86 PMID: 2533575
  2. Control of retroviral RNA splicing through maintenance of suboptimal processing signals.
    Mol Cell Biol. 1990 Feb;10(2):696-704 PMID: 2153921
  3. Mutations in the regions of the Rous sarcoma virus 3' splice sites: implications for regulation of alternative splicing.
    J Virol. 1991 May;65(5):2640-6 PMID: 1850037
  4. Rev is necessary for translation but not cytoplasmic accumulation of HIV-1 vif, vpr, and env/vpu 2 RNAs.
    Genes Dev. 1991 May;5(5):808-19 PMID: 1827422
  5. Primary structure of the human splicing factor ASF reveals similarities with Drosophila regulators.
    Cell. 1991 Jul 26;66(2):373-82 PMID: 1855257
  6. 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
  7. Characterization of Rous sarcoma virus intronic sequences that negatively regulate splicing.
    Virology. 1991 Nov;185(1):99-108 PMID: 1656608
  8. Regulation of human immunodeficiency virus replication.
    Annu Rev Microbiol. 1991;45:219-50 PMID: 1741615
  9. Distinct RNA sequences in the gag region of human immunodeficiency virus type 1 decrease RNA stability and inhibit expression in the absence of Rev protein.
    J Virol. 1992 Jan;66(1):150-9 PMID: 1727477
  10. Intronic sequences and 3' splice sites control Rous sarcoma virus RNA splicing.
    J Virol. 1992 Jan;66(1):6-11 PMID: 1309264
  11. An ATP-independent complex commits pre-mRNA to the mammalian spliceosome assembly pathway.
    Genes Dev. 1991 Dec;5(12B):2534-46 PMID: 1836445
  12. SR proteins: a conserved family of pre-mRNA splicing factors.
    Genes Dev. 1992 May;6(5):837-47 PMID: 1577277
  13. 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
  14. The basic domain of Rev from human immunodeficiency virus type 1 specifically blocks the entry of U4/U6.U5 small nuclear ribonucleoprotein in spliceosome assembly.
    J Virol. 1993 Aug;67(8):4769-76 PMID: 8331728
  15. Rare scleroderma autoantibodies to the U11 small nuclear ribonucleoprotein and to the trimethylguanosine cap of U small nuclear RNAs.
    Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6781-5 PMID: 8341699
  16. Mutation of an RSV intronic element abolishes both U11/U12 snRNP binding and negative regulation of splicing.
    Genes Dev. 1993 Oct;7(10):1926-36 PMID: 8405999
  17. Rev and the fate of pre-mRNA in the nucleus: implications for the regulation of RNA processing in eukaryotes.
    Mol Cell Biol. 1993 Oct;13(10):6180-9 PMID: 8105371
  18. Pathways for selection of 5' splice sites by U1 snRNPs and SF2/ASF.
    EMBO J. 1993 Sep;12(9):3607-17 PMID: 8253084
  19. General splicing factor SF2/ASF promotes alternative splicing by binding to an exonic splicing enhancer.
    Genes Dev. 1993 Dec;7(12B):2598-608 PMID: 8276242
  20. Equine infectious anemia virus trans-regulatory protein Rev controls viral mRNA stability, accumulation, and alternative splicing.
    J Virol. 1994 May;68(5):3102-11 PMID: 8151775
  21. The human splicing factor ASF/SF2 can specifically recognize pre-mRNA 5' splice sites.
    Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3363-7 PMID: 7512732
  22. Presence of negative and positive cis-acting RNA splicing elements within and flanking the first tat coding exon of human immunodeficiency virus type 1.
    Mol Cell Biol. 1994 Jun;14(6):3960-70 PMID: 8196635
  23. Conserved sequences in a class of rare eukaryotic nuclear introns with non-consensus splice sites.
    J Mol Biol. 1994 Jun 10;239(3):357-65 PMID: 8201617
  24. SR proteins promote the first specific recognition of Pre-mRNA and are present together with the U1 small nuclear ribonucleoprotein particle in a general splicing enhancer complex.
    Mol Cell Biol. 1994 Nov;14(11):7670-82 PMID: 7935481
  25. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  26. The human splicing factors ASF/SF2 and SC35 possess distinct, functionally significant RNA binding specificities.
    EMBO J. 1995 Jul 17;14(14):3540-51 PMID: 7543047
  27. SWAP pre-mRNA splicing regulators are a novel, ancient protein family sharing a highly conserved sequence motif with the prp21 family of constitutive splicing proteins.
    Nucleic Acids Res. 1994 Oct 25;22(21):4510-9 PMID: 7971282
  28. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  29. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  30. Nucleotide sequence of Rous sarcoma virus.
    Cell. 1983 Mar;32(3):853-69 PMID: 6299578
  31. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing.
    Gene. 1984 Jun;28(3):351-9 PMID: 6235151
  32. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  33. Rous sarcoma virus nucleic acid-binding protein p12 is necessary for viral 70S RNA dimer formation and packaging.
    J Virol. 1986 Nov;60(2):450-9 PMID: 2430109
  34. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
    Science. 1988 Jan 29;239(4839):487-91 PMID: 2448875
  35. cis-acting intron mutations that affect the efficiency of avian retroviral RNA splicing: implication for mechanisms of control.
    J Virol. 1988 Aug;62(8):2686-95 PMID: 2839694
  36. Regulation of Rous sarcoma virus RNA splicing and stability.
    Mol Cell Biol. 1988 Nov;8(11):4858-67 PMID: 2850470
  37. Synthesis and processing of avian sarcoma retrovirus RNA.
    Adv Virus Res. 1988;35:1-38 PMID: 2852891
  38. Multiple regions in the Rous sarcoma virus src gene intron act in cis to affect the accumulation of unspliced RNA.
    J Virol. 1989 Apr;63(4):1669-76 PMID: 2538650
  39. A suboptimal src 3' splice site is necessary for efficient replication of Rous sarcoma virus.
    Virology. 1995 Feb 1;206(2):1099-107 PMID: 7856084
  40. Inhibition of RNA splicing at the Rous sarcoma virus src 3' splice site is mediated by an interaction between a negative cis element and a chicken embryo fibroblast nuclear factor.
    J Virol. 1995 Aug;69(8):5068-76 PMID: 7609076
  41. Identification of positive and negative splicing regulatory elements within the terminal tat-rev exon of human immunodeficiency virus type 1.
    Mol Cell Biol. 1995 Aug;15(8):4597-605 PMID: 7623851
  42. Presence of exon splicing silencers within human immunodeficiency virus type 1 tat exon 2 and tat-rev exon 3: evidence for inhibition mediated by cellular factors.
    Mol Cell Biol. 1995 Aug;15(8):4606-15 PMID: 7623852
  43. Overexpression of the SR proteins ASF/SF2 and SC35 influences alternative splicing in vivo in diverse ways.
    RNA. 1995 May;1(3):335-46 PMID: 7489505
  44. The superfamily of arginine/serine-rich splicing factors.
    RNA. 1995 Sep;1(7):663-80 PMID: 7585252
  45. The role of branchpoint and 3'-exon sequences in the control of balanced splicing of avian retrovirus RNA.
    Genes Dev. 1991 Feb;5(2):211-20 PMID: 1847346
Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.70.2.1163-1172.1996
Published
1996-02-00
Pages
1163-72
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC189925
Subset
IM
Grants
NCI NIH HHS · CA48746 · United States
External Links
PubMed source DOI: 10.1128/JVI.70.2.1163-1172.1996 PMC Full Text View journal details
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