Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1996 Dec 1;24(23):4684–4692. doi: 10.1093/nar/24.23.4684

B-lineage regulated polyadenylation occurs on weak poly(A) sites regardless of sequence composition at the cleavage and downstream regions.

S A Matis 1, K Martincic 1, C Milcarek 1
PMCID: PMC146313  PMID: 8972854

Abstract

Early/memory and plasma B-cell lines and fibroblasts were analyzed for their ability to use a 5' proximal (variant) versus a 3' distal (constant) poly(A) site, in the absence of a competing splice, from a set of related constructs. The proximal:distal poly(A) site use (P:D ratio) of the resulting cytoplasmic poly(A)+ mRNA is a measure of poly(A) site strength. In this context the immunoglobulin gamma2b secretory-specific poly(A) site showed a P:D ratio of 1:1 in plasma cells, 0.43:1 in early/memory B-cells and an intermediate value in fibroblasts. Meanwhile, a construct with a proximal SV40 early-like poly(A) site produced mRNA with a P:D ratio of >>50:1 in all cell types. Alterations in the region downstream of the proximal poly(A) addition site and at the site itself resulted in changes in the P:D ratio. However, these poly(A) sites, all with a P:D ratio of < or = 5:1, were used most efficiently in plasma cells. Constructs totally devoid of immunoglobulin sequences, but containing heterologous poly(A) sites producing mRNA with P:D ratios of < or = 5:1, were also used more efficiently in plasma cells. We therefore conclude that weak poly(A) sites, regardless of sequence composition, are used more efficiently in plasma cells than in the other cell types.

Full Text

The Full Text of this article is available as a PDF (411.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alt F. W., Bothwell A. L., Knapp M., Siden E., Mather E., Koshland M., Baltimore D. Synthesis of secreted and membrane-bound immunoglobulin mu heavy chains is directed by mRNAs that differ at their 3' ends. Cell. 1980 Jun;20(2):293–301. doi: 10.1016/0092-8674(80)90615-7. [DOI] [PubMed] [Google Scholar]
  2. Brown S. L., Morrison S. L. Developmental regulation of membrane and secretory Ig gamma 2b mRNA. J Immunol. 1989 Mar 15;142(6):2072–2080. [PubMed] [Google Scholar]
  3. Cushley W., Coupar B. E., Mickelson C. A., Williamson A. R. A common mechanism for the synthesis of membrane and secreted immunoglobulin alpha, gamma and mu chains. Nature. 1982 Jul 1;298(5869):77–79. doi: 10.1038/298077a0. [DOI] [PubMed] [Google Scholar]
  4. Denome R. M., Cole C. N. Patterns of polyadenylation site selection in gene constructs containing multiple polyadenylation signals. Mol Cell Biol. 1988 Nov;8(11):4829–4839. doi: 10.1128/mcb.8.11.4829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Edwalds-Gilbert G., Milcarek C. Regulation of poly(A) site use during mouse B-cell development involves a change in the binding of a general polyadenylation factor in a B-cell stage-specific manner. Mol Cell Biol. 1995 Nov;15(11):6420–6429. doi: 10.1128/mcb.15.11.6420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edwalds-Gilbert G., Milcarek C. The binding of a subunit of the general polyadenylation factor cleavage-polyadenylation specificity factor (CPSF) to polyadenylation sites changes during B cell development. Nucleic Acids Symp Ser. 1995;(33):229–233. [PubMed] [Google Scholar]
  7. Flaspohler J. A., Boczkowski D., Hall B. L., Milcarek C. The 3'-untranslated region of membrane exon 2 from the gamma 2a immunoglobulin gene contributes to efficient transcription termination. J Biol Chem. 1995 May 19;270(20):11903–11911. doi: 10.1074/jbc.270.20.11903. [DOI] [PubMed] [Google Scholar]
  8. Flaspohler J. A., Milcarek C. Myelomas and lymphomas expressing the Ig gamma 2a H chain gene have similar transcription termination regions. J Immunol. 1990 Apr 1;144(7):2802–2810. [PubMed] [Google Scholar]
  9. Galli G., Guise J. W., McDevitt M. A., Tucker P. W., Nevins J. R. Relative position and strengths of poly(A) sites as well as transcription termination are critical to membrane versus secreted mu-chain expression during B-cell development. Genes Dev. 1987 Jul;1(5):471–481. doi: 10.1101/gad.1.5.471. [DOI] [PubMed] [Google Scholar]
  10. Genovese C., Harrold S., Milcarek C. Differential mRNA stabilities affect mRNA levels in mutant mouse myeloma cells. Somat Cell Mol Genet. 1991 Jan;17(1):69–81. doi: 10.1007/BF01233206. [DOI] [PubMed] [Google Scholar]
  11. Hall B., Milcarek C. Sequence and polyadenylation site determination of the murine immunoglobulin gamma 2a membrane 3' untranslated region. Mol Immunol. 1989 Sep;26(9):819–826. doi: 10.1016/0161-5890(89)90137-5. [DOI] [PubMed] [Google Scholar]
  12. Hart R. P., McDevitt M. A., Ali H., Nevins J. R. Definition of essential sequences and functional equivalence of elements downstream of the adenovirus E2A and the early simian virus 40 polyadenylation sites. Mol Cell Biol. 1985 Nov;5(11):2975–2983. doi: 10.1128/mcb.5.11.2975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kobrin B. J., Milcarek C., Morrison S. L. Sequences near the 3' secretion-specific polyadenylation site influence levels of secretion-specific and membrane-specific IgG2b mRNA in myeloma cells. Mol Cell Biol. 1986 May;6(5):1687–1697. doi: 10.1128/mcb.6.5.1687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lassman C. R., Matis S., Hall B. L., Toppmeyer D. L., Milcarek C. Plasma cell-regulated polyadenylation at the Ig gamma 2b secretion-specific poly(A) site. J Immunol. 1992 Feb 15;148(4):1251–1260. [PubMed] [Google Scholar]
  15. Lassman C. R., Milcarek C. Regulated expression of the mouse gamma 2b Ig H chain gene is influenced by polyA site order and strength. J Immunol. 1992 Apr 15;148(8):2578–2585. [PubMed] [Google Scholar]
  16. Lebman D. A., Park M. J., Fatica R., Zhang Z. Regulation of usage of membrane and secreted 3' termini of alpha mRNA differs from mu mRNA. J Immunol. 1992 May 15;148(10):3282–3289. [PubMed] [Google Scholar]
  17. Lou H., Gagel R. F., Berget S. M. An intron enhancer recognized by splicing factors activates polyadenylation. Genes Dev. 1996 Jan 15;10(2):208–219. doi: 10.1101/gad.10.2.208. [DOI] [PubMed] [Google Scholar]
  18. Lutz C. S., Murthy K. G., Schek N., O'Connor J. P., Manley J. L., Alwine J. C. Interaction between the U1 snRNP-A protein and the 160-kD subunit of cleavage-polyadenylation specificity factor increases polyadenylation efficiency in vitro. Genes Dev. 1996 Feb 1;10(3):325–337. doi: 10.1101/gad.10.3.325. [DOI] [PubMed] [Google Scholar]
  19. McDevitt M. A., Hart R. P., Wong W. W., Nevins J. R. Sequences capable of restoring poly(A) site function define two distinct downstream elements. EMBO J. 1986 Nov;5(11):2907–2913. doi: 10.1002/j.1460-2075.1986.tb04586.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Milcarek C., Suda-Hartman M., Croll S. C. Changes in abundance of IgG 2a mRNA in the nucleus and cytoplasm of a murine B-lymphoma before and after fusion to a myeloma cell. Mol Immunol. 1996 May-Jun;33(7-8):691–701. doi: 10.1016/0161-5890(96)00009-0. [DOI] [PubMed] [Google Scholar]
  21. Nishikura K., Vuocolo G. A. Synthesis of two mRNAs by utilization of alternate polyadenylation sites: expression of SV40-mouse immunoglobulin mu chain gene recombinants in Cos monkey cells. EMBO J. 1984 Apr;3(4):689–699. doi: 10.1002/j.1460-2075.1984.tb01871.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Niwa M., MacDonald C. C., Berget S. M. Are vertebrate exons scanned during splice-site selection? Nature. 1992 Nov 19;360(6401):277–280. doi: 10.1038/360277a0. [DOI] [PubMed] [Google Scholar]
  23. Oi V. T., Morrison S. L., Herzenberg L. A., Berg P. Immunoglobulin gene expression in transformed lymphoid cells. Proc Natl Acad Sci U S A. 1983 Feb;80(3):825–829. doi: 10.1073/pnas.80.3.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Peterson M. L., Gimmi E. R., Perry R. P. The developmentally regulated shift from membrane to secreted mu mRNA production is accompanied by an increase in cleavage-polyadenylation efficiency but no measurable change in splicing efficiency. Mol Cell Biol. 1991 Apr;11(4):2324–2327. doi: 10.1128/mcb.11.4.2324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Peterson M. L. Regulated immunoglobulin (Ig) RNA processing does not require specific cis-acting sequences: non-Ig RNA can be alternatively processed in B cells and plasma cells. Mol Cell Biol. 1994 Dec;14(12):7891–7898. doi: 10.1128/mcb.14.12.7891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Seipelt R. L., Peterson M. L. Alternative processing of IgA pre-mRNA responds like IgM to alterations in the efficiency of the competing splice and cleavage-polyadenylation reactions. Mol Immunol. 1995 Mar;32(4):277–285. doi: 10.1016/0161-5890(94)00141-m. [DOI] [PubMed] [Google Scholar]
  27. Tsurushita N., Korn L. J. Effects of intron length on differential processing of mouse mu heavy-chain mRNA. Mol Cell Biol. 1987 Jul;7(7):2602–2605. doi: 10.1128/mcb.7.7.2602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Whitelaw E., Proudfoot N. Alpha-thalassaemia caused by a poly(A) site mutation reveals that transcriptional termination is linked to 3' end processing in the human alpha 2 globin gene. EMBO J. 1986 Nov;5(11):2915–2922. doi: 10.1002/j.1460-2075.1986.tb04587.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yan D. H., Weiss E. A., Nevins J. R. Identification of an activity in B-cell extracts that selectively impairs the formation of an immunoglobulin mu s poly(A) site processing complex. Mol Cell Biol. 1995 Apr;15(4):1901–1906. doi: 10.1128/mcb.15.4.1901. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES