Regulation of nuclear poly(A) addition controls the expression of immunoglobulin M secretory mRNA

Phillips, Chris; Jung, Stephen P.; Gunderson, Samuel

doi:10.1093/emboj/20.22.6443

Cited by 50 publications

(74 citation statements)

References 52 publications

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“…The full-length cDNAs of HMGA1a, HMGA1b, and U1-70K were amplified by RT-PCR with attached BamHI and EcoRI sites, and subcloned into pcDNA3(+) vector (Invitrogen) between corresponding restriction enzyme sites. U1-A expression plasmid 37 was a generous gift from Dr S Gunderson.…”

Section: Methodsmentioning

confidence: 99%

Induced HMGA1a expression causes aberrant splicing of Presenilin-2 pre-mRNA in sporadic Alzheimer's disease

et al. 2003

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The aberrant splicing isoform (PS2V), generated by exon 5 skipping of the Presenilin-2 (PS2) gene transcript, is a diagnostic feature of sporadic Alzheimer's disease (AD). We found PS2V is hypoxia-inducible in human neuroblastoma SK-N-SH cells. We purified a responsible trans-acting factor based on its binding to an exon 5 fragment. The factor was identified as the high mobility group A1a protein (HMGA1a; formerly HMG-I). HMGA1a bound to a specific sequence on exon 5, located upstream of the 5 0 splice site. HMGA1a expression was induced by hypoxia and the protein was accumulated in the nuclear speckles with the endogenous splicing factor SC35. Overexpression of HMGA1a generated PS2V, but PS2V was repressed by cotransfection with the U1 snRNP 70K protein that has a strong affinity to HMGA1a. HMGA1a could interfere with U1 snRNP binding to the 5 0 splice site and caused exon 5 skipping. HMGA1a levels were significantly increased in the brain tissue from sporadic AD patients. We propose a novel mechanism of sporadic AD that involves HMGA1a-induced aberrant splicing of PS2 premRNA in the absence of any mutations.

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Section: Methodsmentioning

confidence: 99%

Induced HMGA1a expression causes aberrant splicing of Presenilin-2 pre-mRNA in sporadic Alzheimer's disease

et al. 2003

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“…The forward primers were as follows: for V1S1 and 2, CCGGAATTCCGGACCGTGGACAAGTggtgaGGTAAACCCtgtgTGTACAATGTCTCC-CTGATCATGTCTG; for V1S1, CCGGAATTCCGGACCGTGGACAAGTggtgacGGTAAACCCACACTGT ACAATGTCTCC; and for V1S2, CCGGAATTCCGGACCGTGGACAAGTCCACTGGGTAAACCCtgtgTGTAC-AATGTCTCCCTGATCATGTCTG. The reverse primer was: GCGTC-TAGATAGGG TGGAGGCAAGTATGC as previously described (13).…”

Section: Methodsmentioning

confidence: 99%

“…We have previously shown that the change in stability is regulated by the addition of a poly(A) tail to the secretory mRNA (13). The U1A protein binds upstream of the secretory poly(A) site and inhibits polyadenylation of the secretory mRNA in a developmentally regulated manner.…”

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confidence: 99%

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Sequences Adjacent to the 5′ Splice Site Control U1A Binding Upstream of the IgM Heavy Chain Secretory Poly(A) Site

Phillips

Gunderson

2003

Journal of Biological Chemistry

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We have recently shown that the stability of the alternatively expressed immunoglobulin M heavy chain secretory mRNA is developmentally regulated by U1A. U1A binds novel non-consensus sites upstream of the secretory poly(A) site and inhibits poly(A) tail addition in undifferentiated cells. U1A's dependence for binding and function upon a stem-loop structure has been extensively characterized for the consensus sites. We therefore probed the structure surrounding the novel U1A binding sites. We show that two of the three novel binding sites represent the major single-stranded regions upstream of the secretory poly(A) site, consistent with a major role at this site. The strength of binding and ability of U1A to inhibit poly(A) polymerase correlate with the accessibility of the novel sites. However, long range interactions are responsible for maintaining them in an open configuration. Mutation of an RNase V1-sensitive site 102 nucleotides upstream, directly adjacent to the competing 5 splice site, changes the structure of one the U1A binding sites and thus abolishes the binding of the second U1A molecule and the ability of U1A ability to inhibit poly(A) polymerase activity at this site. These sites bind U1A via its N-terminal domain but with a 10-fold lower affinity than U1 small nuclear RNA. This lower binding affinity is more conducive to U1A's regulation of poly(A) tail addition to heterologous mRNA.The major source of diversity in metazoans is the ability to process pre-mRNA into alternative mRNAs via the selection of alternative splice sites and alternative 3Ј-ends, giving rise to a range of proteins encoded by the same gene that are differentially expressed during growth and development (for review see Ref.

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“…The level of a specific mRNA transcript within a cell is regulated by either the stability of the transcript and/or the rate at which the transcript is produced (49). To determine whether the stability of mature IgG1 transcript was affected by stimulation of CD86 and/or ␤ 2 AR, actinomycin D was added to cultured B cells on day 6 after activation.…”

Section: Stability Of Mature Igg1 Transcript Following Stimulation Ofmentioning

confidence: 99%

Selective Regulation of Mature IgG1 Transcription by CD86 and β2-Adrenergic Receptor Stimulation

Podojil

Sanders

2003

The Journal of Immunology

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Stimulation of CD86 and the β2-adrenergic receptor (β2AR) on a B cell, either alone or together, is known to increase the level of IgG1 protein produced by a CD40 ligand/IL-4-activated B cell. It is also known that the mechanism by which CD40 and IL-4R stimulation on a B cell increases the level of IgG1 protein is by increasing germline γ1 transcription, IgG1 class switching, and mature IgG1 transcription, while the molecular mechanism responsible for mediating the CD86- and β2AR-induced effect remains unknown. In the present study using real-time PCR we show that the level of mature IgG1 transcription increases in CD40 ligand/IL-4-activated B cells following stimulation of either CD86 and/or β2AR, and that this increase reflects the increase in IgG1 protein. Furthermore, we show that the CD86- and/or β2AR-induced increase in mature IgG1 transcript is due to an increase in the rate of mature IgG1 transcription, as determined by nuclear run-on analysis. This effect is additive when both receptors are stimulated and is lost when B cells from CD86- and β2AR-deficient mice are used. In contrast, the level of germline γ1 transcription, the stability of mature IgG1 transcript, the number of IgG1-positive B cells, and the number of IgG1-secreting B cells did not change. These results provide the first evidence that CD86 and/or β2AR stimulation on a CD40 ligand/IL-4-activated B cell increases the level of IgG1 protein produced per cell by increasing the rate of mature IgG1 transcription.

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Regulation of nuclear poly(A) addition controls the expression of immunoglobulin M secretory mRNA

Cited by 50 publications

References 52 publications

Induced HMGA1a expression causes aberrant splicing of Presenilin-2 pre-mRNA in sporadic Alzheimer's disease

Induced HMGA1a expression causes aberrant splicing of Presenilin-2 pre-mRNA in sporadic Alzheimer's disease

Sequences Adjacent to the 5′ Splice Site Control U1A Binding Upstream of the IgM Heavy Chain Secretory Poly(A) Site

Selective Regulation of Mature IgG1 Transcription by CD86 and β2-Adrenergic Receptor Stimulation

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