Multiple features contribute to efficient constitutive splicing of an unusually large exon

Bruce, Shirley R.; Peterson, Martha L.

doi:10.1093/nar/29.11.2292

Cited by 15 publications

(15 citation statements)

References 39 publications

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“…Our data are consistent with B cells splicing weaker splice sites more efficiently than plasma cells; the Cµ4 5Ј splice site is known to be suboptimal (Peterson and Perry 1989), the cryptic 5Ј splice site in pIgR exon 4 is weaker than the authentic pIgR 5Ј splice site (Bruce and Peterson 2001), and the µAV 3Ј splice site is suboptimal and requires a downstream ESE. This enhanced splice activity in B cells, in combination with lower cleavage-polyadenylation activity, would result in Cµ4-M1 splicing competing successfully with cleavage-polyadenylation at the suboptimal µs poly(A) site in B cells.…”

Section: Discussionsupporting

confidence: 86%

“…3C). We also monitored expression by RT-PCR (data not shown), which provides information on the size of the pIgR exon spliced into the D d RNA, as described previously (Bruce et al 1999;Bruce and Peterson 2001). The ratio of full-length to cryptic RNA expressed in the S194 plasma-cell line was similar to that seen previously in HepG2 cells; in pIgR, exon 4 was spliced intact, whereas the 5ЈSS mutation partially activated the previously identified upstream cryptic 5Ј splice site (Fig.…”

Section: The Splicing Environment Of B Cells and Plasma Cells Is Diffsupporting

confidence: 70%

“…For each construct, the ratio of full-length to cryptically spliced RNA was 7-to 11-fold higher in the plasma cells than in the B cells. Several other pIgR mutations (Bruce and Peterson 2001) were also expressed in M12 and S194 cells with similar results; in all cases, the upstream cryptic splice site was recognized more frequently in the M12 B cells than in the S194 plasma cells (data not shown). These results were also confirmed by stably transfecting pIgR and 5ЈSS into the M12 and S194 cell lines (data not shown).…”

Section: The Splicing Environment Of B Cells and Plasma Cells Is Diffsupporting

confidence: 64%

“…We have shown previously that the unusually large fourth exon from the mouse polymeric Ig receptor (pIgR) gene, when placed in the third intron of the mouse major histocom- Fig. 2A), is constitutively spliced into the mRNA in the human HepG2 liver-cell line (Bruce et al 1999;Bruce and Peterson 2001). However, when the 5Ј splice site of pIgR exon 4 was weakened by a point mutation (D d -pIgR 5ЈSS or 5ЈSS, Fig.…”

Section: The Splicing Environment Of B Cells and Plasma Cells Is Diffmentioning

confidence: 99%

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

Bruce¹,

Dingle²,

Peterson³

2003

RNA

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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 membraneassociated 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.

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Section: Discussionsupporting

confidence: 86%

Section: The Splicing Environment Of B Cells and Plasma Cells Is Diffsupporting

confidence: 70%

Section: The Splicing Environment Of B Cells and Plasma Cells Is Diffsupporting

confidence: 64%

Section: The Splicing Environment Of B Cells and Plasma Cells Is Diffmentioning

confidence: 99%

See 2 more Smart Citations

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

Bruce¹,

Dingle²,

Peterson³

2003

RNA

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“…Constitutive splicing can be performed by the minimal spliceosome and involves ligation of the distal end of the upstream exon with the nearest splice acceptor, which is at the proximal end of the next exon in the sequence. Among the different mechanisms of constitutive and alternative splicing, the possible exclusion of sequences where both the 5Ј and the 3Ј intronic junctions are located within an exon represents a further pattern of alternative splicing (33). In alternative splicing, one or more splice acceptors and associated exons are skipped, resulting in removal of their coding information from the final mRNA.…”

Section: Discussionmentioning

confidence: 99%

Serine-Arginine-rich Protein p30 Directs Alternative Splicing of Glucocorticoid Receptor Pre-mRNA to Glucocorticoid Receptor β in Neutrophils

Leung

Kisich

2003

Journal of Biological Chemistry

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Glucocorticoid (GC) insensitivity is a major clinical challenge in the treatment of many inflammatory diseases. It has been shown previously that GC insensitivity, in several inflammatory cell types, is due to an overabundance of the ␤ isoform of the glucocorticoid receptor (GCR␤) relative to the ligand binding isoform, GCR␣. GCR␤ functions as a dominant inhibitor of GCR␣ action. A number of GCR isoforms are created from the same pre-mRNA transcript via alternative splicing, and the factor or factors that control alternative splicing of GCR pre-mRNA are of great importance. In the current study, we have identified the predominant alternative splicing factor present in human neutrophils, which is known to be exceptionally GC-insensitive. The predominant alternative splicing factor in neutrophils is SRp30c, which is one of several highly conserved serinearginine-rich (SR) proteins that are involved in both constitutive and alternative splicing in eukaryotic cells. Inhibition of SRp30c expression with antisense oligonucleotide strongly inhibited expression of GCR␤ and stimulated expression of GCR␣. Antisense molecules targeted to other SR proteins had no effect. Our data indicate that SRp30c is necessary for alternative splicing of the GCR pre-mRNA to create mRNA encoding GCR␤.Glucocorticoid (GC) 1 insensitivity is a major clinical challenge in the treatment of chronic inflammatory diseases. The pharmacologic actions of GCs are mediated through intracellular receptors, the glucocorticoid receptors (GCR). There are two isoforms of GCR in human cells, GCR␣ and GCR␤, which are generated from a single gene via alternative splicing of the primary RNA transcript. Several studies indicate that GC insensitivity has been associated with increased expression of GCR␤ (1-7). GCR␤ is truncated at the C terminus, which corresponds to the ligand binding domain. Thus, GCR␤ cannot bind GC. In addition, GCR␤ does not transactivate GC-sensitive genes and functions as a dominant inhibitor of GCR␣ (8). Previous studies suggest that GCR␣:GCR␤ heterodimer formation may account for the reduced effectiveness of GC action in cells overexpressing GCR␤ (8). Recent experiments in our lab demonstrated that overexpression of human GCR␤ by mouse hybridoma cells results in the development of GC insensitivity by these cells (9).Regulation of GCR␤ expression is not well understood. Different cell types from the same individual can have very different ratios of GCR␣ to GCR␤. For example, freshly isolated peripheral blood neutrophils are GC-insensitive. Both the absolute level of GCR␤ and the ratio of GCR␤ to GCR␣ protein are much higher in neutrophils than in peripheral blood mononuclear cells (PBMC) from the same individuals (10). The ratio of GCR␤ to GCR␣ can be altered by cytokine stimulation. After stimulation of neutrophils with IL-8, GCR␤ mRNA levels increased remarkably, and GCR␣ mRNA decreased to undetectable levels. Similarly, exposure of PBMC to IL-2 and IL-4 resulted in increased GCR␤ expression and development of steroid insensitivity (1,...

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Immunoglobulin heavy chain gene regulation through polyadenylation and splicing competition

Peterson

2010

WIREs RNA

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The immunoglobulin heavy chain (IgH) genes, which encode one of the two chains of antibody molecules, were the first cellular genes shown to undergo developmentally regulated alternative RNA processing. These genes produce two different mRNAs from a single primary transcript. One mRNA is cleaved and polyadenylated at an upstream poly(A) signal while the other mRNA removes this poly(A) signal by RNA splicing and is cleaved and polyadenylated at a downstream poly(A) site. A broad range of studies have been performed to understand the mechanism of IgH RNA processing regulation during B lymphocyte development. The model that has emerged is much more complex than envisioned by the earliest view of regulation through poly(A) signal choice. Regulation requires that the IgH gene contain competing splice and cleavage-polyadenylation reactions with balanced efficiencies. Because non-IgH genes with these structural features also can be regulated, IgH gene-specific sequence elements are not required for regulation. Changes in cleavage-polyadenylation and RNA splicing, as well as pol II elongation, all contribute to IgH developmental RNA processing regulation. Multiple factors are likely involved in the regulation during B lymphocyte maturation. Additional biologically relevant factors that contribute to IgH regulation remain to be identified and incorporated into a mechanistic model for regulation. Much of the work to date confirms the complex nature of IgH mRNA regulation and suggests that a thorough understanding of this control will remain a challenge. However, it is also likely that such understanding will help elucidate novel mechanisms of RNA processing regulation.

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Multiple features contribute to efficient constitutive splicing of an unusually large exon

Cited by 15 publications

References 39 publications

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

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

Serine-Arginine-rich Protein p30 Directs Alternative Splicing of Glucocorticoid Receptor Pre-mRNA to Glucocorticoid Receptor β in Neutrophils

Immunoglobulin heavy chain gene regulation through polyadenylation and splicing competition

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