“…Tau4R and Tau3R isoforms differ in their binding affinity to microtubules (Lee et al 1989; Goedert and Jakes 1990; Goode et al 2000; Utton et al 2002; Tomoo et al 2005; Konzack et al 2007; LeBoeuf et al 2008). In the adult human brain, alternative splicing of tau exon 10 is tightly regulated to maintain the ratio of Tau4R to Tau3R at approximately one, whereas tau is expressed as almost exclusively Tau3R isoform in the fetal brain (Neve et al 1986; Lichtenberg-Kraag and Mandelkow 1990; Andreadis et al 1992; Gao et al 2000). Disruption of this delicate balance in the Tau4R/ Tau3R ratio is associated with tauopathies (Hutton et al 1998; Hasegawa et al 1998; Grover et al 1999; Hasegawa et al 1999; Spillantini et al 2000; Connell et al 2001; Cairns et al 2007; Mackenzie et al 2010).…”
Microtubule binding protein Tau has been implicated in a wide range of neurodegenerative disorders collectively classified as tauopathies. Exon 10 of the human tau gene, which codes for a microtubule binding repeat region, is alternatively spliced to form Tau protein isoforms containing either four or three microtubule binding repeats, Tau4R and Tau3R, respectively. The levels of different Tau splicing isoforms are fine-tuned by alternative splicing with the ratio of Tau4R/Tau3R maintained approximately at one in adult neurons. Mutations that disrupt tau exon 10 splicing regulation cause an imbalance of different tau splicing isoforms and have been associated with tauopathy. To search for factors interacting with tau pre-messenger RNA (pre-mRNA) and regulating tau exon 10 alternative splicing, we performed a yeast RNA–protein interaction screen and identified polypyrimidine tract binding protein associated splicing factor (PSF) as a candidate tau exon 10 splicing regulator. UV crosslinking experiments show that PSF binds to the stem-loop structure at the 5′ splice site downstream of tau exon 10. This PSF-interacting RNA element is distinct from known PSF binding sites previously identified in other genes. Overexpression of PSF promotes tau exon 10 exclusion, whereas down-regulation of the endogenous PSF facilitates exon 10 inclusion. Immunostaining shows that PSF is expressed in the human brain regions affected by tauopathy. Our data reveal a new player in tau exon 10 alternative splicing regulation and uncover a previously unknown mechanism of PSF in regulating tau pre-mRNA splicing.
“…Tau4R and Tau3R isoforms differ in their binding affinity to microtubules (Lee et al 1989; Goedert and Jakes 1990; Goode et al 2000; Utton et al 2002; Tomoo et al 2005; Konzack et al 2007; LeBoeuf et al 2008). In the adult human brain, alternative splicing of tau exon 10 is tightly regulated to maintain the ratio of Tau4R to Tau3R at approximately one, whereas tau is expressed as almost exclusively Tau3R isoform in the fetal brain (Neve et al 1986; Lichtenberg-Kraag and Mandelkow 1990; Andreadis et al 1992; Gao et al 2000). Disruption of this delicate balance in the Tau4R/ Tau3R ratio is associated with tauopathies (Hutton et al 1998; Hasegawa et al 1998; Grover et al 1999; Hasegawa et al 1999; Spillantini et al 2000; Connell et al 2001; Cairns et al 2007; Mackenzie et al 2010).…”
Microtubule binding protein Tau has been implicated in a wide range of neurodegenerative disorders collectively classified as tauopathies. Exon 10 of the human tau gene, which codes for a microtubule binding repeat region, is alternatively spliced to form Tau protein isoforms containing either four or three microtubule binding repeats, Tau4R and Tau3R, respectively. The levels of different Tau splicing isoforms are fine-tuned by alternative splicing with the ratio of Tau4R/Tau3R maintained approximately at one in adult neurons. Mutations that disrupt tau exon 10 splicing regulation cause an imbalance of different tau splicing isoforms and have been associated with tauopathy. To search for factors interacting with tau pre-messenger RNA (pre-mRNA) and regulating tau exon 10 alternative splicing, we performed a yeast RNA–protein interaction screen and identified polypyrimidine tract binding protein associated splicing factor (PSF) as a candidate tau exon 10 splicing regulator. UV crosslinking experiments show that PSF binds to the stem-loop structure at the 5′ splice site downstream of tau exon 10. This PSF-interacting RNA element is distinct from known PSF binding sites previously identified in other genes. Overexpression of PSF promotes tau exon 10 exclusion, whereas down-regulation of the endogenous PSF facilitates exon 10 inclusion. Immunostaining shows that PSF is expressed in the human brain regions affected by tauopathy. Our data reveal a new player in tau exon 10 alternative splicing regulation and uncover a previously unknown mechanism of PSF in regulating tau pre-mRNA splicing.
“…Tau4R and Tau3R show distinct activities in microtubule binding (43,68,69,70,(100)(101)(102)(103)105). tau exon 10 alternative splicing undergoes developmental stage-specific regulation, with almost exclusive expression of Tau3R in the fetal brain and both isoforms being expressed equally (Tau4R/Tau3R ratio of 1:1) in the adult human brain (3,4,38,60,74,88,90).…”
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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