TDP43 depletion rescues aberrant CFTR exon 9 skipping

Ayala, Youhna M.; Pagani, Franco; Baralle, Francisco E.

doi:10.1016/j.febslet.2006.01.052

Cited by 78 publications

(86 citation statements)

References 19 publications

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“…This mechanism is supported by the lack of change in Cdk6 levels upon TDP-43 depletion in chicken cells, which express a Cdk6 transcript devoid of GU repeats. The involvement of TDP-43 in the control of RNA processing has been seen in other systems where GU repeats are involved, i.e., CFTR and ApoAII alternative splicing (2)(3)(4)(5). The elucidation of the precise step of RNA processing controlled by TDP-43 in the case of Cdk6 needs further investigation.…”

Section: Discussionmentioning

confidence: 99%

“…TDP-43 was depleted from HeLa cells by RNAi routinely achieving Ͼ90% silencing as measured by Western blot, 48 h after small interfering RNA (siRNA) transfection (3,5). RNA microarray analysis was performed on TDP-43 depleted and control treated cells.…”

Section: Tdp-43 Down-regulation Alters the Expression Of Prb-related mentioning

confidence: 99%

“…Members of the hnRNP family serve multiple roles in the generation and processing of RNA, including transcription, splicing, transport, and stability. TDP-43 inhibits exon recognition during splicing upon recruitment to the 3Ј splice site of the cystic fibrosis transmembrane conductance regulator (CFTR) and apolipoprotein AII transcripts via a sequence of GU repeats (2)(3)(4)(5). The binding affinity of the recombinant human, worm, and fly homologues for this target sequence is remarkably high, measured in the low nanomolar range (6).…”

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confidence: 99%

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TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression

Ayala

Misteli

Baralle

2008

Proc. Natl. Acad. Sci. U.S.A.

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214

197

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T DP-43 belongs to the family of heterogeneous nuclear ribonucleoproteins (hnRNPs) and binds single-stranded RNA via its N-terminal RNA recognition motif (1). Members of the hnRNP family serve multiple roles in the generation and processing of RNA, including transcription, splicing, transport, and stability. TDP-43 inhibits exon recognition during splicing upon recruitment to the 3Ј splice site of the cystic fibrosis transmembrane conductance regulator (CFTR) and apolipoprotein AII transcripts via a sequence of GU repeats (2-5). The binding affinity of the recombinant human, worm, and fly homologues for this target sequence is remarkably high, measured in the low nanomolar range (6). TDP-43 also has been implicated in the transcription regulation of HIV and the spermatid-specific gene SP-10 through promoter association (7,8). More recently, TDP-43 was identified as the main ubiquitinated component of cytoplasmic inclusions in neurodegenerative diseases, specifically frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS) (9, 10). Abnormal aggregation of TDP-43 in the cytoplasm now is thought to define a class of frontotemporal dementias termed TDP-43 proteinopathies. Mislocalization and the consequent loss of TDP-43 function in neuronal cells may represent a common event in FTLD pathogenesis. Despite the increasing awareness of processes involving TDP-43, the cellular role of the protein still is poorly defined.We depleted TDP-43 by RNA interference (RNAi) to identify TDP-43-regulated transcripts. Our results point to cyclindependent kinase 6 (Cdk6) as a unique target of TDP-43 regulation and suggest that TDP-43 inhibits Cdk6 expression through recruitment to the GU-rich transcript. Simultaneously, we found that TDP-43 silencing alters cell cycle distribution and induces apoptosis. Table 1 lists 16 of these proteins whose functions have been associated with retinoblastoma protein (pRb) activity. The tumor suppressor pRb is essential for the control of cell cycle progression, cellular differentiation, and maintenance of genome integrity. Inactivation of pRb occurs through its gradual phosphorylation by Cdks during the G 1 phase of the cell division cycle resulting in the activation of transcription factors that promote cell proliferation and enable transition on to the S phase (see ref. 11 for review). Our RNA microarray analyses showed altered levels of transcripts coding for proteins whose functions are related to the control of cell cycle progression (Cdk6, POLD4, cyclin B1, Cdk2, UBE2C, and SKP2). In addition, some of these factors are known to either directly interact with pRb (e.g., HDAC1, RBBP4, and CRI1), or act in response to pRb modulation (e.g., E2F8 and NAP1L1) (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22).

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Section: Discussionmentioning

confidence: 99%

Section: Tdp-43 Down-regulation Alters the Expression Of Prb-related mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression

Ayala

Misteli

Baralle

2008

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

214

197

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“…The siRNA sequences were as follows: TDP-43 siRNA 5 0 -GCAAAGCCAAGAUGAGCCU-3 0 , Luciferase siRNA 5 0 -UAAGGCUAUGAAGAGAUAC-3 0 . 43,44 RNA preparation, synthesis of cDNA, PCRs RNA preparation. Total cellular RNA was isolated with TRI REAGENT (Sigma) following manufacturer's instructions.…”

Section: Cell Culture Transient Transfectionsmentioning

confidence: 99%

TDP-43 regulatesβ-adducin(Add2) transcript stability

et al. 2014

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Keywords: Add2, DSE, mRNA stability, TDP-43, 3 0 end processing, 3 0 UTR Abbreviations: RBP, RNA binding protein, DSE, downstream element, PAS, polyadenylation site, ActD, actinomycin D.TDP-43 is an RNA-binding protein involved in several steps of mRNA metabolism including transcription, splicing and stability. It is also involved in ALS and FTD, neurodegenerative diseases characterized by TDP-43 nuclear depletion. We previously identified TDP-43 as a binder of the downstream element (DSE) of the b-Adducin (Add2) brain-specific polyadenylation site (A4 PAS), suggesting its involvement in pre-mRNA 3 0 end processing. Here, by using chimeric minigenes, we showed that TDP-43 depletion in HeLa and HEK293 cells resulted in down-regulation of both the chimeric and endogenous Add2 transcripts. Despite having confirmed TDP-43-DSE in vitro interaction, we demonstrated that the in vivo effect was not mediated by the TDP-43-DSE interaction. In fact, substitution of the Add2 DSE with viral E-SV40 and L-SV40 DSEs, which are not TDP-43 targets, still resulted in decreased Add2 mRNA levels after TDP-43 downregulation. In addition, we failed to show interaction between TDP-43 and key polyadenylation factors, such as CstF-64 and CPSF160 and excluded TDP-43 involvement in pre-mRNA cleavage and regulation of polyA tail length. These evidences allowed us to exclude the pre-hypothesized role of TDP43 in modulating 3 0 end processing of Add2 pre-mRNA. Finally, we showed that TDP-43 regulates Add2 gene expression levels by increasing Add2 mRNA stability. Considering that Add2 in brain participates in synapse assembly, synaptic plasticity and their stability, and its genetic inactivation in mice leads to LTP, LTD, learning and motor-coordination deficits, we hypothesize that a possible loss of Add2 function by TDP-43 depletion may contribute to ALS and FTD disease states.

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“…Under physiological conditions, functions of TDP-43 are associated with various pathways by binding to DNA, RNA or other nuclear proteins (Buratti et al, 2001;Buratti et al, 2004;Ayala et al, 2005;Mercado et al, 2005). It was reported that TDP-43 can recognize the intronic UG tract of CFTR pre-mRNA, thusly promoting skipping of exon 9 in CFTR mRNA (Wang et al, 2004;Ayala et al, 2006;Buratti et al, 2008), suggesting its role in RNA splicing regulation. TDP-43 also can interact with many other splicing related protein Freibaum et al, 2010) via the C-terminal glycine-rich domain of the protein (Wang et al, 2004;Ayala et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Staurosporine Induced Apoptosis Rapidly Downregulates TDP-43 in Glioma Cells

Nan

Zhu²,

Tan³

et al. 2014

Asian Pacific Journal of Cancer Prevention

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TDP-43 is a ubiquitously expressed DNA/RNA binding protein that has recently attracted attention for its involvement in neurodegenerative diseases. While TDP-43 has been found to participate in various important cellular activities including stress and apoptosis, little is known about its role in cancer cells. Here we report that staurosporine (STS) induced apoptosis in U87 glioma cells is associated with rapid downregulation of TDP-43 at both mRNA and protein levels. The latter is dependent on activation of caspase 3. More importantly, we have shown that knockdown of TDP-43 by specific siRNA dramatically enhanced cytotoxicity of STS. These results suggest that normal level of TDP-43 may be protective for cancer cells under apoptotic insult.

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TDP43 depletion rescues aberrant CFTR exon 9 skipping

Cited by 78 publications

References 19 publications

TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression

TDP-43 regulates retinoblastoma protein phosphorylation through the repression of cyclin-dependent kinase 6 expression

TDP-43 regulatesβ-adducin(Add2) transcript stability

Staurosporine Induced Apoptosis Rapidly Downregulates TDP-43 in Glioma Cells

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