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PMID: 3013627 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A single nucleotide difference at the 3' end of an intron causes differential splicing of two histocompatibility genes.

The EMBO journal ·Vol. 5 ·No. 5 ·1986-05-00 ·Pages 957-65

Archibald AL, Thompson NA, Kvist S

Abstract

The murine histocompatibility class I genes, H-2 Kb and Kk, display considerable homology at their 3' ends. In fact, from exon 5 to the termination codon, only two nucleotides differ between the two genes, one at the 5' end and the other at the 3' end of intron 7. Despite this similarity, the gene products have distinctly different mol. wts as determined by SDS-PAGE. By constructing two hybrid genes, pC2 and pC4, we demonstrated that it is the cytoplasmic parts of the antigens (encoded by exons 6-8) which are responsible for the major difference in mol. wt. We have used site-directed mutagenesis to change the two nucleotides in intron 7 of the H-2 Kk gene to those present in the H-2 Kb gene. S1 nuclease mapping has been used to identify the actual splice site of the authentic Kb and Kk genes, the hybrid genes and the mutagenized genes. We have shown that it is the 3' nucleotide difference, nine nucleotides upstream of the 3' splice site, which causes the different excision of intron 7 of the Kb gene. The 5' nucleotide difference does not alter the splicing. The choice of branch points and 3' splice signals for intron 7 of five H-2 class I genes, is discussed.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cell Line DNA Restriction Enzymes Electrophoresis, Polyacrylamide Gel Genes H-2 Antigens/genetics,isolation & purification Major Histocompatibility Complex Mice Molecular Weight RNA Splicing Sequence Homology, Nucleic Acid
Chemicals
H-2 Antigens DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Archibald A L
Thompson N A
Kvist S
References (60)
60 references, click to expand
  1. Mouse histocompatibility genes: structure and organisation of a Kd gene.
    EMBO J. 1983;2(2):245-54 PMID: 11894934
  2. An adenovirus type 2 glycoprotein blocks cell surface expression of human histocompatibility class I antigens.
    Cell. 1985 Jul;41(3):987-97 PMID: 3924414
  3. Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences.
    Cell. 1985 Aug;42(1):355-67 PMID: 3160483
  4. On the origin of RNA splicing and introns.
    Cell. 1985 Sep;42(2):397-400 PMID: 2411416
  5. Role of the 3' splice site consensus sequence in mammalian pre-mRNA splicing.
    Nature. 1985 Oct 24-30;317(6039):732-4 PMID: 4058579
  6. Exonuclease III: use for DNA sequence analysis and in specific deletions of nucleotides.
    Methods Enzymol. 1983;100:60-96 PMID: 6312265
  7. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  8. Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4853-7 PMID: 283395
  9. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373-6 PMID: 286319
  10. MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness.
    Adv Immunol. 1979;27:51-177 PMID: 92183
  11. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  12. Fine specificity analysis with monoclonal antibodies of antigens controlled by the major histocompatibility complex and by the Qa/TL region in mice.
    Immunol Rev. 1979;47:175-206 PMID: 398326
  13. Directed deletion of a yeast transfer RNA intervening sequence.
    Science. 1980 Sep 19;209(4463):1396-400 PMID: 6997991
  14. Alternatives for splicing: recognizing the ends of introns.
    Cell. 1980 Nov;22(2 Pt 2):324-6 PMID: 6160915
  15. Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression.
    J Immunol. 1981 Jan;126(1):317-21 PMID: 6935293
  16. A subcloning strategy for DNA sequence analysis.
    Nucleic Acids Res. 1980 Dec 11;8(23):5541-9 PMID: 6258146
  17. Structure of C-terminal half of two H-2 antigens from cloned mRNA.
    Nature. 1981 Jul 2;292(5818):78-81 PMID: 6895103
  18. Oligonucleotide directed mutagenesis of the human beta-globin gene: a general method for producing specific point mutations in cloned DNA.
    Nucleic Acids Res. 1981 Aug 11;9(15):3647-56 PMID: 7279669
  19. DNA sequence of a gene encoding a BALB/c mouse Ld transplantation antigen.
    Science. 1982 Feb 5;215(4533):679-82 PMID: 7058332
  20. A catalogue of splice junction sequences.
    Nucleic Acids Res. 1982 Jan 22;10(2):459-72 PMID: 7063411
  21. Isolation of a cDNA clone for the murine transplantation antigen H-2Kb.
    Proc Natl Acad Sci U S A. 1982 May;79(10):3270-4 PMID: 6954478
  22. Membrane insertion and oligomeric assembly of HLA-DR histocompatibility antigens.
    Cell. 1982 May;29(1):61-9 PMID: 6955026
  23. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.
    J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
  24. Abnormal RNA splicing causes one form of alpha thalassemia.
    Cell. 1982 Jul;29(3):895-902 PMID: 7151175
  25. A single-base change at a splice site in a beta 0-thalassemic gene causes abnormal RNA splicing.
    Cell. 1982 Jul;29(3):903-11 PMID: 7151176
  26. Unusual splice sites revealed by mutagenic inactivation of an authentic splice site of the rabbit beta-globin gene.
    Nature. 1983 Jan 6;301(5895):38-43 PMID: 6296682
  27. A cDNA clone containing the entire coding sequence of a mouse H-2Kd histocompatibility antigen.
    Nucleic Acids Res. 1983 Mar 11;11(5):1567-77 PMID: 6298749
  28. Human beta-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei.
    Cell. 1983 Mar;32(3):681-94 PMID: 6550524
  29. Specific transcription and RNA splicing defects in five cloned beta-thalassaemia genes.
    Nature. 1983 Apr 14;302(5909):591-6 PMID: 6188062
  30. pEMBL: a new family of single stranded plasmids.
    Nucleic Acids Res. 1983 Mar 25;11(6):1645-55 PMID: 6300771
  31. Polyadenylylated nuclear RNA contains branches.
    Proc Natl Acad Sci U S A. 1983 Feb;80(4):950-4 PMID: 6573664
  32. Studies on transformation of Escherichia coli with plasmids.
    J Mol Biol. 1983 Jun 5;166(4):557-80 PMID: 6345791
  33. Splicing of adenovirus RNA in a cell-free transcription system.
    Proc Natl Acad Sci U S A. 1983 Sep;80(17):5230-4 PMID: 6577417
  34. Evidence for the biochemical role of an internal sequence in yeast nuclear mRNA introns: implications for U1 RNA and metazoan mRNA splicing.
    Cell. 1983 Sep;34(2):395-403 PMID: 6616616
  35. Expression and function of transplantation antigens with altered or deleted cytoplasmic domains.
    Cell. 1983 Sep;34(2):535-44 PMID: 6604582
  36. Splicing of in vitro synthesized messenger RNA precursors in HeLa cell extracts.
    Cell. 1983 Nov;35(1):89-99 PMID: 6194902
  37. Genes of the major histocompatibility complex in mouse and man.
    Science. 1983 Nov 18;222(4625):727-33 PMID: 6356354
  38. Alternative RNA splicing in expression of the H-2K gene.
    Nature. 1983 Dec 8-14;306(5943):602-4 PMID: 6689056
  39. Construction, expression and recognition of an H-2 molecule lacking its carboxyl terminus.
    Nature. 1984 Feb 2-8;307(5950):432-6 PMID: 6363940
  40. Point mutations identify the conserved, intron-contained TACTAAC box as an essential splicing signal sequence in yeast.
    Cell. 1984 Mar;36(3):645-53 PMID: 6365330
  41. Normal and mutant human beta-globin pre-mRNAs are faithfully and efficiently spliced in vitro.
    Cell. 1984 Apr;36(4):993-1005 PMID: 6323033
  42. Messenger RNA splicing in vitro: an excised intervening sequence and a potential intermediate.
    Cell. 1984 Jun;37(2):415-27 PMID: 6722880
  43. Identification of an H-2Kd gene using a specific cDNA probe.
    EMBO J. 1982;1(4):467-71 PMID: 6329684
  44. A minimal intron length but no specific internal sequence is required for splicing the large rabbit beta-globin intron.
    Cell. 1984 Jul;37(3):915-25 PMID: 6204770
  45. Synthesis and characterization of an octanucleotide containing the EcoRI recognition sequence with a phosphorothioate group at the cleavage site.
    Biochemistry. 1984 Jul 17;23(15):3443-53 PMID: 6087894
  46. Excision of an intact intron as a novel lariat structure during pre-mRNA splicing in vitro.
    Cell. 1984 Aug;38(1):317-31 PMID: 6088074
  47. Cytolytic T cells recognize the two amino-terminal domains of H-2 K antigens in tandem in influenza A infected cells.
    Cell. 1984 Aug;38(1):79-87 PMID: 6088078
  48. Molecular biology. Excision of introns in lariat form.
    Nature. 1984 Sep 13-19;311(5982):103-4 PMID: 6472468
  49. Lariat RNA's as intermediates and products in the splicing of messenger RNA precursors.
    Science. 1984 Aug 31;225(4665):898-903 PMID: 6206566
  50. Intron splicing: a conserved internal signal in introns of animal pre-mRNAs.
    Proc Natl Acad Sci U S A. 1984 Dec;81(23):7417-20 PMID: 6209716
  51. The RNA lariat: a new ring to the splicing of mRNA precursors.
    Cell. 1984 Dec;39(3 Pt 2):423-5 PMID: 6568879
  52. In vivo splicing products of the rabbit beta-globin pre-mRNA.
    Cell. 1984 Dec;39(3 Pt 2):589-602 PMID: 6096012
  53. In vivo characterization of yeast mRNA processing intermediates.
    Cell. 1984 Dec;39(3 Pt 2):603-10 PMID: 6096013
  54. Lariat structures are in vivo intermediates in yeast pre-mRNA splicing.
    Cell. 1984 Dec;39(3 Pt 2):611-21 PMID: 6096014
  55. Complete nucleotide sequence of the murine H-2Kk gene. Comparison of three H-2K locus alleles.
    Nucleic Acids Res. 1984 Dec 21;12(24):9473-87 PMID: 6096831
  56. Characterization of the branch site in lariat RNAs produced by splicing of mRNA precursors.
    Nature. 1985 Feb 14-20;313(6003):552-7 PMID: 2578627
  57. DNA sequence of the mouse H-2Dd transplantation antigen gene.
    Proc Natl Acad Sci U S A. 1985 Feb;82(4):1175-9 PMID: 3856254
  58. Intron sequences involved in lariat formation during pre-mRNA splicing.
    Cell. 1985 May;41(1):95-105 PMID: 3888410
  59. Cryptic branch point activation allows accurate in vitro splicing of human beta-globin intron mutants.
    Cell. 1985 Jul;41(3):833-44 PMID: 3879973
  60. The DNA sequence of the H-2kb gene: evidence for gene conversion as a mechanism for the generation of polymorphism in histocompatibilty antigens.
    EMBO J. 1983;2(3):453-62 PMID: 11894963
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1986-05-00
Pages
957-65
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1166888
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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