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RNA (2003), 9:1264-1273. Published by Cold Spring Harbor Laboratory Press. Copyright © 2003 RNA Society

B-cell and plasma-cell splicing differences: A potential role in regulated immunoglobulin RNA processing

SHIRLEY R. BRUCE1,3, R.W. CAMERON DINGLE1 and MARTHA L. PETERSON1,2

1 Department of Microbiology, Immunology, and Molecular Genetics, Department of Pathology and Laboratory Medicine, and
2 The Lucille Parker Markey Cancer Center, University of Kentucky College of Medicine, Lexington, Kentucky 40536, USA

Reprint requests to: Martha L. Peterson, Department of Microbiology, Immunology, and Molecular Genetics, Department of Pathology and Laboratory Medicine, and The Lucille Parker Markey Cancer Center, University of Kentucky College of Medicine, Lexington, Kentucky 40536, USA; e-mail: mlpete01{at}uky.edu; fax: (859) 323-2094.

The immunoglobulin µ pre-mRNA is alternatively processed at its 3' end by competing splice and cleavage-polyadenylation reactions to generate mRNAs encoding the membrane-associated or secreted forms of the IgM protein, respectively. The relative use of the competing processing pathways varies during B-lymphocyte development, and it has been established previously that cleavage-polyadenylation activity is higher in plasma cells, which secrete IgM, than in B cells, which produce membrane-associated IgM. To determine whether RNA-splicing activity varies during B-lymphocyte development to contribute to µ RNA-processing regulation, we first demonstrate that µ pre-mRNA processing is sensitive to artificial changes in the splice environment by coexpressing SR proteins with the µ gene. To explore differences between the splice environments of B cells and plasma cells, we analyzed the splicing patterns from two different chimeric non-Ig genes that can be alternatively spliced but have no competing cleavage-polyadenylation reaction. The ratio of intact exon splicing to cryptic splice site use from one chimeric gene differs between several B-cell and several plasma-cell lines. Also, the amount of spliced RNA is higher in B-cell than plasma-cell lines from a set of genes whose splicing is dependent on a functional exonic splice enhancer. Thus, there is clear difference between the B-cell and plasma-cell splicing environments. We propose that both general cleavage-polyadenylation and general splice activities are modulated during B-lymphocyte development to ensure proper regulation of the alternative µ RNA processing pathways.

Keywords: Cleavage-polyadenylation; RNA splicing; SR proteins


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