Friend of Prmt1, FOP is a Novel Component of the Nuclear SMN Complex Isolated Using Biotin Affinity Purification

Izumikawa, Keiichi; Ishikawa, Hideaki; Yoshikawa, Harunori; Terukina, Goro; Miyazawa, Naoki; Nakayama, Hiroshi; Nobe, Yuko; Taoka, Masato; Yamauchi, Yoshio; Philipsen, Sjaak; Isobe, Toshiaki; Takahashi, Nobuhiro

doi:10.4172/jpb.s7-002

Cited by 6 publications

(17 citation statements)

References 42 publications

(64 reference statements)

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“…DAP-Chtop and DAP-Lyar (a negative control) were pulled down with 100 μl of Ni-NTA (Qiagen) in 8 ml xTractor buffer (Clontech) containing 1% (v/v) Empigen-BB (SIGMA) from T-Rex 293 cells (1.0 × 10 8 cells) expressing DAP-Chtop (9), and DAP-Lyar (10) after induction with doxycycline for 24 h, respectively. The pulled down DAP-tagged proteins were purified further with 100 μl of FLAG-M2 agarose (SIGMA) using a solution containing 50 mM Tris-HCl pH7.4, 150 mM NaCl, 0.5% IGEPAL CA-630 and 1% Empigen-BB, dialyzed in buffer D (20 mM Hepes (pH7.9), 100 mM KCl, 0.2 mM EDTA, 20% Glycerol, 1 mM DTT) for over 12 h, and used for in vitro splicing assay.…”

Section: Methodsmentioning

confidence: 99%

“…(9). A Biotinylated (BT)-oligonucleotide (5′-GACTATATTGCAAGTCGTCACGGC-3′) probe was used to detect U4 snRNA (12).…”

Section: Methodsmentioning

confidence: 99%

“…BT-RNA (1 pmol) was added to the extract and subjected to immunoprecipitation using anti-FLAG magnetic beads as described in Supplementary Data. RNA isolated from FLAG-tagged protein complexes was subjected to denaturing urea-PAGE and Northern blotting (9,12). BT-RNA was detected with stabilized Streptavidin-HRP Conjugate (Thermo Scientific, 89880D).…”

Section: Methodsmentioning

confidence: 99%

“…Finally, Chtop is a component of the nuclear SMN (survival of motor neuron protein) complex that contains U5 and U6 but lacks U1 and U2 small nuclear RNAs (snRNAs) (9). Chtop is required for association of the SMN complex with heterogeneous nuclear RNPs (hnRNPs), histones and various RNA-binding proteins (9).…”

Section: Introductionmentioning

confidence: 99%

“…Chtop is required for association of the SMN complex with heterogeneous nuclear RNPs (hnRNPs), histones and various RNA-binding proteins (9). Despite accumulating evidence that Chtop is involved in various aspects of gene expression, the underlying mechanism by which the expression level of Chtop itself is regulated remains undetermined.…”

Section: Introductionmentioning

confidence: 99%

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Chtop (Chromatin target of Prmt1) auto-regulates its expression level via intron retention and nonsense-mediated decay of its own mRNA

et al. 2016

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Chtop (chromatin target of Prmt1) regulates various aspects of gene expression including transcription and mRNA export. Despite these important functions, the regulatory mechanism underlying Chtop expression remains undetermined. Using Chtop-expressing human cell lines, we demonstrate that Chtop expression is controlled via an autoregulatory negative feedback loop whereby Chtop binds its own mRNA to retain intron 2 during splicing; a premature termination codon present at the 5′ end of intron 2 leads to nonsense-mediated decay of the mRNA. We also show that Chtop interacts with exon 2 of Chtop mRNA via its arginine-glycine-rich (RG) domain, and with intron 2 via its N-terminal (N1) domain; both are required for retention of intron 2. In addition, we show that hnRNP H accelerates intron 2 splicing of Chtop mRNA in a manner dependent on Chtop expression level, suggesting that Chtop and hnRNP H regulate intron 2 retention of Chtop mRNA antagonistically. Thus, the present study provides a novel molecular mechanism by which mRNA and protein levels are constitutively regulated by intron retention.

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Section: Methodsmentioning

confidence: 99%

“…(9). A Biotinylated (BT)-oligonucleotide (5′-GACTATATTGCAAGTCGTCACGGC-3′) probe was used to detect U4 snRNA (12).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Chtop (Chromatin target of Prmt1) auto-regulates its expression level via intron retention and nonsense-mediated decay of its own mRNA

et al. 2016

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Arginine methylation: the promise of a ‘silver bullet’ for brain tumours?

et al. 2021

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Despite intense research efforts, our pharmaceutical repertoire against high-grade brain tumours has not been able to increase patient survival for a decade and life expectancy remains at less than 16 months after diagnosis, on average. Inhibitors of protein arginine methyltransferases (PRMTs) have been developed and investigated over the past 15 years and have now entered oncology clinical trials, including for brain tumours. This review collates recent advances in the understanding of the role of PRMTs and arginine methylation in brain tumours. We provide an up-to-date literature review on the mechanisms for PRMT regulation. These include endogenous modulators such as alternative splicing, miRNA, post-translational modifications and PRMT–protein interactions, and synthetic inhibitors. We discuss the relevance of PRMTs in brain tumours with a particular focus on PRMT1, -2, -5 and -8. Finally, we include a future perspective where we discuss possible routes for further research on arginine methylation and on the use of PRMT inhibitors in the context of brain tumours.

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TDP-43 stabilises the processing intermediates of mitochondrial transcripts

Izumikawa

Nobe

Yoshikawa

et al. 2017

Sci Rep

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The 43-kDa trans-activating response region DNA-binding protein 43 (TDP-43) is a product of a causative gene for amyotrophic lateral sclerosis (ALS). Despite of accumulating evidence that mitochondrial dysfunction underlies the pathogenesis of TDP-43–related ALS, the roles of wild-type TDP-43 in mitochondria are unknown. Here, we show that the small TDP-43 population present in mitochondria binds directly to a subset of mitochondrial tRNAs and precursor RNA encoded in L-strand mtDNA. Upregulated expression of TDP-43 stabilised the processing intermediates of mitochondrial polycistronic transcripts and their products including the components of electron transport and 16S mt-rRNA, similar to the phenotype observed in cells deficient for mitochondrial RNase P. Conversely, TDP-43 deficiency reduced the population of processing intermediates and impaired mitochondrial function. We propose that TDP-43 has a novel role in maintaining mitochondrial homeostasis by regulating the processing of mitochondrial transcripts.

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Friend of Prmt1, FOP is a Novel Component of the Nuclear SMN Complex Isolated Using Biotin Affinity Purification

Cited by 6 publications

References 42 publications

Chtop (Chromatin target of Prmt1) auto-regulates its expression level via intron retention and nonsense-mediated decay of its own mRNA

Chtop (Chromatin target of Prmt1) auto-regulates its expression level via intron retention and nonsense-mediated decay of its own mRNA

Arginine methylation: the promise of a ‘silver bullet’ for brain tumours?

TDP-43 stabilises the processing intermediates of mitochondrial transcripts

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