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PMID: 2878362 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Sequences near the 3' secretion-specific polyadenylation site influence levels of secretion-specific and membrane-specific IgG2b mRNA in myeloma cells.

Molecular and cellular biology ·Vol. 6 ·No. 5 ·1986-05-00 ·Pages 1687-97

Kobrin BJ, Milcarek C, Morrison SL

Abstract

The expressed immunoglobulin gamma 2b (IgG2b) heavy-chain gene of 4T001 was cloned into the shuttle vector pSV2-gpt and transfected into myeloma J558L and lymphoma A20.2J. Northern blots indicated that the transfected gamma 2b gene was processed in a manner similar to the endogenous heavy chain in both lymphoma and myeloma cells. To identify sequences important for immunoglobulin mRNA processing, we constructed deletions around the secretion-specific polyadenylation site and introduced the deleted genes into J558L cells. The BAL deletion lacked 670 base pairs of intervening sequence between secreted and membrane regions; the Kpn deletion lacked 830 base pairs in this region. J558L cells transfected with either the entire gamma 2b gene or the delta BAL vector produced predominantly secretion-specific gamma 2b mRNA and protein. J558L cells transfected with the delta Kpn vector produced approximately equimolar amounts of secretion-specific and membrane-specific gamma 2b mRNA. Both 55,000-dalton secreted and 62,000-dalton putative surface IgG2b proteins were detected in the delta Kpn transfectants. We conclude that sequences absent in the Kpn deletion but present in the BAL deletion exert an important role in the production of secretion-specific mRNA. The Kpn deletion removes the normal site of cleavage and poly(A) addition, and it is possible that it is the absence of this site which changes the processing pattern. Alternatively, it is possible that sequences absent in the Kpn deletion but present in the BAL deletion function in regulating the production of predominantly secretion-specific mRNA in myeloma cells. The possible role of a highly conserved sequence found in this region is discussed.

MeSH Terms
Animals Base Sequence Cell Line DNA Restriction Enzymes Genes Immunoglobulin G/genetics Lymphoma/immunology Mice Plasmacytoma/immunology Plasmids Poly A/genetics RNA, Messenger/genetics,metabolism
Chemicals
Immunoglobulin G RNA, Messenger Poly A DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kobrin B J
Milcarek C
Morrison S L
References (50)
50 references, click to expand
  1. Cell-specific expression of secreted versus membrane forms of immunoglobulin gamma 2b mRNA involves selective use of alternate polyadenylation sites.
    Mol Cell Biol. 1985 Oct;5(10):2514-20 PMID: 3939252
  2. Immunoglobulin messenger RNAs in murine cell lines that have characteristics of immature B lymphocytes.
    Cell. 1979 Dec;18(4):1333-9 PMID: 117905
  3. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
  4. Complete nucleotide sequence of mouse immunoglobulin mu gene and comparison with other immunoglobulin heavy chain genes.
    Nucleic Acids Res. 1980 Sep 11;8(17):3933-45 PMID: 6255422
  5. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs.
    Cell. 1981 Apr;24(1):251-60 PMID: 6113054
  6. Organization and expression of immunoglobulin genes in fetal liver hybridomas.
    Proc Natl Acad Sci U S A. 1981 Jan;78(1):247-51 PMID: 6787586
  7. Comparison of mouse immunoglobulin gamma 2a and gamma 2b chain genes suggests that exons can be exchanged between genes in a multigenic family.
    Proc Natl Acad Sci U S A. 1981 Apr;78(4):2442-6 PMID: 6787604
  8. Roles of surface-bound immunoglobulin molecules in regulating the replication and maturation to immunoglobulin secretion of B lymphocytes.
    Immunol Rev. 1980;52:89-114 PMID: 7021398
  9. Expression of surface and secreted IgG2a by a murine B-lymphoma before and after hybridization to myeloma cells.
    Mol Immunol. 1981 Apr;18(4):311-22 PMID: 6795453
  10. Gene segments encoding transmembrane carboxyl termini of immunoglobulin gamma chains.
    Cell. 1981 Oct;26(1 Pt 1):19-27 PMID: 6799207
  11. On immunoglobulin heavy chain gene switching: two gamma 2b genes are rearranged via switch sequences in MPC-11 cells but only one is expressed.
    Nucleic Acids Res. 1982 Jan 22;10(2):611-30 PMID: 6278424
  12. Mouse IgA heavy chain gene sequence: implications for evolution of immunoglobulin hinge axons.
    Proc Natl Acad Sci U S A. 1981 Dec;78(12):7684-8 PMID: 6801659
  13. Linkage and sequence homology of two human immunoglobulin gamma heavy chain constant region genes.
    Proc Natl Acad Sci U S A. 1982 Mar;79(6):1984-8 PMID: 6804948
  14. mRNA for surface immunoglobulin gamma chains encodes a highly conserved transmembrane sequence and a 28-residue intracellular domain.
    Proc Natl Acad Sci U S A. 1982 Mar;79(6):2008-12 PMID: 6804950
  15. B-cell subpopulations identified by two-colour fluorescence analysis.
    Nature. 1982 Jun 17;297(5867):589-91 PMID: 7045679
  16. Multiple immunoglobulin heavy-chain gene transcripts in Abelson murine leukemia virus-transformed lymphoid cell lines.
    Mol Cell Biol. 1982 Apr;2(4):386-400 PMID: 6810096
  17. The nucleotide sequence of a human immunoglobulin C gamma1 gene.
    Nucleic Acids Res. 1982 Jul 10;10(13):4071-9 PMID: 6287432
  18. The ontogeny and distribution of B cells in normal and mutant immune-defective CBA/N mice: two-parameter analysis of surface IgM and IgD.
    J Immunol. 1983 Feb;130(2):619-25 PMID: 6600246
  19. Isolation and characterization of a variant of mouse plasmacytoma J558 synthesizing a 110,000-dalton immunoglobulin heavy chain and of secondary variants synthesizing either a 55,000-dalton or an 80,000-dalton immunoglobulin heavy chain: possible implications.
    Mol Cell Biol. 1982 Sep;2(9):1134-44 PMID: 6184610
  20. Immunoglobulin gene expression in transformed lymphoid cells.
    Proc Natl Acad Sci U S A. 1983 Feb;80(3):825-9 PMID: 6402777
  21. Localization of murine Igh-1a allotypic determinants by using a panel of mouse myeloma variant immunoglobulins.
    J Immunol. 1983 Apr;130(4):1967-9 PMID: 6187830
  22. Unusual sequences in the murine immunoglobulin mu-delta heavy-chain region.
    Nature. 1983 Dec 1-7;306(5942):483-7 PMID: 6417547
  23. Mode of regulation of immunoglobulin mu- and delta-chain expression varies during B-lymphocyte maturation.
    Cell. 1984 Feb;36(2):329-38 PMID: 6319016
  24. Are U4 small nuclear ribonucleoproteins involved in polyadenylation?
    Nature. 1984 May 10-16;309(5964):179-82 PMID: 6325940
  25. B cell activation: three steps and their variations.
    Cell. 1984 Jul;37(3):713-20 PMID: 6234992
  26. Requirement of a downstream sequence for generation of a poly(A) addition site.
    Cell. 1984 Jul;37(3):993-9 PMID: 6744418
  27. Structural analysis of the murine IgG3 constant region gene.
    EMBO J. 1984 Sep;3(9):2041-6 PMID: 6092053
  28. Sequences on the 3' side of hexanucleotide AAUAAA affect efficiency of cleavage at the polyadenylation site.
    Mol Cell Biol. 1984 Aug;4(8):1460-8 PMID: 6149460
  29. Chimeric human antibody molecules: mouse antigen-binding domains with human constant region domains.
    Proc Natl Acad Sci U S A. 1984 Nov;81(21):6851-5 PMID: 6436822
  30. Role of the conserved AAUAAA sequence: four AAUAAA point mutants prevent messenger RNA 3' end formation.
    Science. 1984 Nov 30;226(4678):1045-51 PMID: 6208611
  31. Calcitonin/calcitonin gene-related peptide transcription unit: tissue-specific expression involves selective use of alternative polyadenylation sites.
    Mol Cell Biol. 1984 Oct;4(10):2151-60 PMID: 6334229
  32. Loss of a consensus splice signal in a mutant immunoglobulin gene eliminates the CH1 domain exon from the mRNA.
    Mol Cell Biol. 1984 Jul;4(7):1270-7 PMID: 6095057
  33. mRNA precursor splicing in vivo: sequence requirements determined by deletion analysis of an intervening sequence.
    Proc Natl Acad Sci U S A. 1985 May;82(9):2885-9 PMID: 3857622
  34. The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini.
    Nucleic Acids Res. 1985 Feb 25;13(4):1347-68 PMID: 2987822
  35. A sequence downstream of A-A-U-A-A-A is required for formation of simian virus 40 late mRNA 3' termini in frog oocytes.
    Proc Natl Acad Sci U S A. 1985 Jun;82(12):3949-53 PMID: 2987956
  36. Identification of sequences in the herpes simplex virus thymidine kinase gene required for efficient processing and polyadenylation.
    Mol Cell Biol. 1985 Aug;5(8):2104-13 PMID: 3018551
  37. Expression and regulation of immunoglobulin heavy chain gene transfected into lymphoid cells.
    EMBO J. 1983;2(8):1373-8 PMID: 10872333
  38. Synthesis, assembly, and secretion of gamma globulin by mouse myeloma cells. I. Adaptation of the Merwin plasma cell tumor-11 to culture, cloning, and characterization of gamma globulin subunits.
    J Exp Med. 1970 Mar 1;131(3):515-41 PMID: 4189836
  39. Demonstration of heavy and light chain antigenic determinants on the cell-bound receptor for antigen. Similarities between membrane-attached and humoral antibodies produced by the same cell.
    J Exp Med. 1970 Dec 1;132(6):1233-49 PMID: 4101973
  40. Differentiation of B lymphocytes from stem cell precursors.
    Adv Exp Med Biol. 1973;29(0):11-8 PMID: 4604328
  41. Cell surface immunoglobulin. XI. The appearance of an IgD-like molecule on murine lymphoid cells during ontogeny.
    J Exp Med. 1975 Jan 1;141(1):206-15 PMID: 1090697
  42. Early production of intracellular IgM by B-lymphocyte precursors in mouse.
    Nature. 1976 Jan 22;259(5540):224-6 PMID: 814466
  43. Immunoglobulin-receptors revisited.
    Science. 1975 Sep 19;189(4207):964-9 PMID: 1083069
  44. Demonstration that IgG memory is carried by IgG-bearing cells.
    Eur J Immunol. 1976 Jul;6(7):467-72 PMID: 1086788
  45. Structure of the adenovirus 2 early mRNAs.
    Cell. 1978 Jul;14(3):695-711 PMID: 688389
  46. The synthesis and processing of the messenger RNAs specifying heavy and light chain immunoglobulins in MPC-11 cells.
    Cell. 1978 Dec;15(4):1495-509 PMID: 103631
  47. Transformation of mammalian cells with genes from procaryotes and eucaryotes.
    Cell. 1979 Apr;16(4):777-85 PMID: 222468
  48. Cloning and complete nucleotide sequence of mouse immunoglobulin gamma 1 chain gene.
    Cell. 1979 Oct;18(2):559-68 PMID: 115593
  49. Sequence of the cloned gene for the constant region of murine gamma 2b immunoglobulin heavy chain.
    Science. 1979 Dec 14;206(4424):1303-6 PMID: 117549
  50. Expression of a bacterial gene in mammalian cells.
    Science. 1980 Sep 19;209(4463):1422-7 PMID: 6251549
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.6.5.1687-1697.1986
Published
1986-05-00
Pages
1687-97
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC367696
Subset
IM
Grants
NIAID NIH HHS · AI 19042 · United States
NCI NIH HHS · CA 16850 · United States
NCI NIH HHS · CA 22376 · United States
External Links
PubMed source DOI: 10.1128/mcb.6.5.1687-1697.1986 PMC Full Text View journal details
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