SR protein 9G8 modulates splicing of tau exon 10 via its proximal downstream intron, a clustering region for frontotemporal dementia mutations

Gao, Lei; Wang, Junning; Wang, Yingzi; Andreadis, Athena

doi:10.1016/j.mcn.2006.10.004

Cited by 52 publications

(74 citation statements)

References 55 publications

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“…One microgram of total RNA was used for first-strand cDNA synthesis with oligo-(dT) [15][16][17][18] by using the Omniscript Reverse Transcription kit (Qiagen). PCR was performed by using PrimeSTAR TM HS DNA Polymerase (Takara Bio Inc., Otsu, Shiga, Japan) with forward primer 5Ј-GGTGTCCACTC-CCAGTTCAA-3Ј and reverse primer 5Ј-CCCTGGTTTA-TGATGGATGTTGCCTAATGAG-3Ј to measure alternative splicing of tau exon 10.…”

Section: Quantitation Of Tau Exon 10 Splicing By Reverse Transcriptiomentioning

confidence: 99%

“…Immune complex was eluted with elution buffer (1% SDS, 0.1 M NaHCO 3 , and 50 units/ml RNasin Plus RNase Inhibitor). The cross-linking was reversed by incubation with 200 mM NaCl at 65°C for at least 2 h. After digestion with 20 mM Tris-HCl, pH 6.5, 0.5 M EDTA, 0.8 mg/ml Proteinase K (Invitrogen), RNA was extracted by the RNeasy Mini kit (Qiagen) and subjected to first-strand cDNA synthesis with oligo-(dT) [15][16][17][18] by using the Omniscript Reverse Transcription kit (Qiagen). cDNA was amplified by PrimeSTAR TM HS DNA Polymerase (Takara Bio Inc.) with two sets of primers against pre-mRNA of tau: primer set 1 (forward (5Ј-AGGC-GGGTCCAGGGTGGCGTGTCACTCATC-3Ј) and reverse (5Ј-CTAATAATTCAAGCCACAGCACGGCGCATGGGACG-3Ј), and primer set 2 (forward (5Ј-ATGCCTTCAGAGCAGC-CCTCTATCC-3Ј) and reverse (5Ј-ACGTGTGAAGGTACT-CACACTGCCG-3Ј)).…”

Section: Quantitation Of Tau Exon 10 Splicing By Reverse Transcriptiomentioning

confidence: 99%

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Cyclic AMP-dependent Protein Kinase Regulates the Alternative Splicing of Tau Exon 10

Shi

Qian

Yin

et al. 2011

Journal of Biological Chemistry

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Hyperphosphorylation and deposition of tau into neurofibrillary tangles is a hallmark of Alzheimer disease (AD).Alternative splicing of tau exon 10 generates tau isoforms containing three or four microtubule binding repeats (3R-tau and 4R-tau), which are equally expressed in adult human brain. Dysregulation of exon 10 causes neurofibrillary degeneration. Here, we report that cyclic AMP-dependent protein kinase, PKA, phosphorylates splicing factor SRSF1, modulates its binding to tau pre-mRNA, and promotes tau exon 10 inclusion in cultured cells and in vivo in rat brain. PKA-C␣, but not PKA-C␤, interacts with SRSF1 and elevates SRSF1-mediated tau exon 10 inclusion. In AD brain, the decreased level of PKA-C␣ correlates with the increased level of 3R-tau. These findings suggest that a downregulation of PKA dysregulates the alternative splicing of tau exon 10 and contributes to neurofibrillary degeneration in AD by causing an imbalance in 3R-tau and 4R-tau expression.Tau is a neuronal microtubule-associated protein, the function of which is to stimulate microtubule assembly and stabilize microtubules. Hyperphosphorylation of tau leads to its aggregation into neurofibrillary tangles, a hallmark of Alzheimer disease (AD) 2 and related neurodegenerative diseases called tauopathies (1-4). Adult human brain expresses six different tau isoforms from a single gene by alternative splicing of exons 2, 3, and 10 of its pre-mRNA (5). The exon 10 encodes the second microtubule binding repeat (6). Alternative splicing of exon 10 generates tau with three or four microtubule binding repeats (3R-tau or 4R-tau), which is under developmental and cell type-specific regulation. Only 3R-tau is expressed during embryogenesis, whereas 3R-tau and 4R-tau are expressed in approximately equal amounts in adult human brain (6, 7). Several mutations in tau gene result in either an increase or a decrease in 4R-tau expression and cause frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), one of the tauopathies (8). Thus, alteration in the 3R-tau/ 4R-tau ratio is sufficient to trigger neurodegeneration in frontotemporal dementia and might also play a role in other neurodegenerative disorders such as Pick's disease, progressive nuclear palsy, or corticobasal degeneration in which the 3R-tau/4R-tau ratio is markedly altered (9 -12). Thus, the regulation of alternative splicing of human tau exon 10 has been of critical interest. However, results of studies of the alternative splicing of tau exon 10 in AD brain have been contradictory (13)(14)(15). Recent studies have shown that aggregation and deposition of 3R-tau may be associated with more advanced stages (16,17).Alternative splicing of tau exon 10 is regulated by several trans-acting factors, including serine-and arginine-rich (SR) proteins, and their phosphorylation (18 -24). Splicing factor 2/alternative splicing factor (ASF/SF2), now named SRSF1 (serine/arginine-rich splicing factor 1) (25), is a prototypical SR protein that participates in both constitutive and alternativ...

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Section: Quantitation Of Tau Exon 10 Splicing By Reverse Transcriptiomentioning

confidence: 99%

Section: Quantitation Of Tau Exon 10 Splicing By Reverse Transcriptiomentioning

confidence: 99%

Cyclic AMP-dependent Protein Kinase Regulates the Alternative Splicing of Tau Exon 10

Shi

Qian

Yin

et al. 2011

Journal of Biological Chemistry

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“…The splicing factors SRp20, SRp30c, SRp55, Tra2-b, and 9G8 have all been shown to affect exon 10 splicing. Each of these proteins contains an arginineserine repeat (RS) domain, characteristic of the SR family of splicing factors (Yu et al 2004;Wang et al 2005;Gao et al 2007). SR proteins can act as either alternative splicing enhancers or repressors (Long and Caceres 2009;Shepard and Hertel 2009).…”

Section: Introductionmentioning

confidence: 99%

The cardiotonic steroid digitoxin regulates alternative splicing through depletion of the splicing factors SRSF3 and TRA2B

Anderson

Lin²,

Xiao³

et al. 2012

RNA

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Modulation of alternative pre-mRNA splicing is a potential approach to therapeutic targeting for a variety of human diseases. We investigated the mechanism by which digitoxin, a member of the cardiotonic steroid class of drugs, regulates alternative splicing. Transcriptome-wide analysis identified a large set of alternative splicing events that change after digitoxin treatment. Within and adjacent to these regulated exons, we identified enrichment of potential binding sites for the splicing factors SRp20 (SRSF3/SFRS3) and Tra2-b (SFRS10/TRA2B). We further find that both of these proteins are depleted from cells by digitoxin treatment. Characterization of SRp20 and Tra2-b splicing targets revealed that many, but not all, digitoxin-induced splicing changes can be attributed to the depletion of one or both of these factors. Re-expression of SRp20 or Tra2-b after digitoxin treatment restores normal splicing of their targets, indicating that the digitoxin effect is directly due to these factors. These results demonstrate that cardiotonic steroids, long prescribed in the clinical treatment of heart failure, have broad effects on the cellular transcriptome through these and likely other RNA binding proteins. The approach described here can be used to identify targets of other potential therapeutics that act as alternative splicing modulators.

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“…In some forms of familial FTLD-tau, splicing mutations disrupt tau exon 10 splicing regulation and alter the Tau4R/Tau3R ratio in the brain, leading to neurodegeneration (17,19,23,24,26,43,49,55,97,110). Several studies have begun to reveal cis-acting regulatory elements and trans-acting factors that regulate tau exon 10 splicing (9,26,27,28,37,45,60,61,67,106). One of the cis-acting elements is a stem-loop structure downstream of the 5Ј splice site of exon 10 (45, 54, 60, 103, 104).…”

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confidence: 99%

RNA Helicase p68 (DDX5) Regulates tau Exon 10 Splicing by Modulating a Stem-Loop Structure at the 5′ Splice Site

Kar

Fushimi

Zhou

et al. 2011

Molecular and Cellular Biology

112

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Regulation of tau exon 10 splicing plays an important role in tauopathy. One of the cis elements regulating tau alternative splicing is a stem-loop structure at the 5 splice site of tau exon 10. The RNA helicase(s) modulating this stem-loop structure was unknown. We searched for splicing regulators interacting with this stem-loop region using an RNA affinity pulldown-coupled mass spectrometry approach and identified DDX5/ RNA helicase p68 as an activator of tau exon 10 splicing. The activity of p68 in stimulating tau exon 10 inclusion is dependent on RBM4, an intronic splicing activator. RNase H cleavage and U1 protection assays suggest that p68 promotes conformational change of the stem-loop structure, thereby increasing the access of U1snRNP to the 5 splice site of tau exon 10. This study reports the first RNA helicase interacting with a stem-loop structure at the splice site and regulating alternative splicing in a helicase-dependent manner. Our work uncovers a previously unknown function of p68 in regulating tau exon 10 splicing. Furthermore, our experiments reveal functional interaction between two splicing activators for tau exon 10, p68 binding at the stem-loop region and RBM4 interacting with the intronic splicing enhancer region.

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SR protein 9G8 modulates splicing of tau exon 10 via its proximal downstream intron, a clustering region for frontotemporal dementia mutations

Cited by 52 publications

References 55 publications

Cyclic AMP-dependent Protein Kinase Regulates the Alternative Splicing of Tau Exon 10

Cyclic AMP-dependent Protein Kinase Regulates the Alternative Splicing of Tau Exon 10

The cardiotonic steroid digitoxin regulates alternative splicing through depletion of the splicing factors SRSF3 and TRA2B

RNA Helicase p68 (DDX5) Regulates tau Exon 10 Splicing by Modulating a Stem-Loop Structure at the 5′ Splice Site

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