Characterization of a naturally occurring truncated Dicer

Mosca, Nicola; Staręga-Rosłan, Julia; Castiello, Filomena; Russo, Andrew F.; Krzyżosiak, Włodzimierz J.; Potenza, Nicoletta

doi:10.1007/s11033-015-3878-6

Cited by 6 publications

(4 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Prior to western blotting for Dicer detection, the beads, after washing, were resuspended in a 10 μl of PBS and 10 μl 3× sodium dodecylsulphate (SDS) sample buffer, heated for 5 min at 95°C and loaded onto a 5% Tris-acetate SDS-page (acrylamide:bis = 49:1) in XT Tricine buffer (Bio-Rad) as previously described ( 41 , 42 ). The blots were probed with anti-Dicer antibody (Cell Signaling Technology, 1:1000) and then HRP-conjugated secondary antibody, anti-rabbit (Sigma, 1:1000).…”

Section: Methodsmentioning

confidence: 99%

Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer

2015

Self Cite

View full text Add to dashboard Cite

The ribonuclease Dicer excises mature miRNAs from a diverse group of precursors (pre-miRNAs), most of which contain various secondary structure motifs in their hairpin stem. In this study, we analyzed Dicer cleavage in hairpin substrates deprived of such motifs. We searched for the factors other than the secondary structure, which may influence the length diversity and heterogeneity of miRNAs. We found that the nucleotide sequence at the Dicer cleavage site influences both of these miRNA characteristics. With regard to cleavage mechanism, we demonstrate that the Dicer RNase IIIA domain that cleaves within the 3′ arm of the pre-miRNA is more sensitive to the nucleotide sequence of its substrate than is the RNase IIIB domain. The RNase IIIA domain avoids releasing miRNAs with G nucleotide and prefers to generate miRNAs with a U nucleotide at the 5′ end. We also propose that the sequence restrictions at the Dicer cleavage site might be the factor that contributes to the generation of miRNA duplexes with 3′ overhangs of atypical lengths. This finding implies that the two RNase III domains forming the single processing center of Dicer may exhibit some degree of flexibility, which allows for the formation of these non-standard 3′ overhangs.

show abstract

Section: Methodsmentioning

confidence: 99%

Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer

2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…T-Dicer was detectable in 29 of 55 primary neuroblastic tumours, but not in the normal and other cancer tissues tested. Additionally, t-Dicer functions less efficiently than full length DICER, but not due to dominant negative effects [142]. Further pre-clinical studies are required to ascertain if DROSHA, DICER, and their variants are therapeutic targets or biomarkers in neuroblastoma.…”

Section: Other Rbps Reported In Neuroblastoma Involved In Mirna Biogementioning

confidence: 99%

Identification of RNA-Binding Proteins as Targetable Putative Oncogenes in Neuroblastoma

Bell

Hagemann

Holien

et al. 2020

IJMS

View full text Add to dashboard Cite

Neuroblastoma is a common childhood cancer with almost a third of those affected still dying, thus new therapeutic strategies need to be explored. Current experimental therapies focus mostly on inhibiting oncogenic transcription factor signalling. Although LIN28B, DICER and other RNA-binding proteins (RBPs) have reported roles in neuroblastoma development and patient outcome, the role of RBPs in neuroblastoma is relatively unstudied. In order to elucidate novel RBPs involved in MYCN-amplified and other high-risk neuroblastoma subtypes, we performed differential mRNA expression analysis of RBPs in a large primary tumour cohort (n = 498). Additionally, we found via Kaplan–Meier scanning analysis that 685 of the 1483 tested RBPs have prognostic value in neuroblastoma. For the top putative oncogenic candidates, we analysed their expression in neuroblastoma cell lines, as well as summarised their characteristics and existence of chemical inhibitors. Moreover, to help explain their association with neuroblastoma subtypes, we reviewed candidate RBPs’ potential as biomarkers, and their mechanistic roles in neuronal and cancer contexts. We found several highly significant RBPs including RPL22L1, RNASEH2A, PTRH2, MRPL11 and AFF2, which remain uncharacterised in neuroblastoma. Although not all RBPs appear suitable for drug design, or carry prognostic significance, we show that several RBPs have strong rationale for inhibition and mechanistic studies, representing an alternative, but nonetheless promising therapeutic strategy in neuroblastoma treatment.

show abstract

“…On the basis of previous reports, the apoptotic machinery appears to be implicated in the regulation of Dicer. Active caspase-3 cleaves Dicer into a truncated form and truncated Dicer (tDicer) triggers cell death (Mosca et al, 2015). Western blotting analysis was performed to determine whether the decrease in Dicer levels Figure 3 Activation of caspase-1 during hypoxia-induced apoptosis of T98G cells.…”

Section: Dicer Is Downregulated During Hypoxia and Knockdown Of Dicermentioning

confidence: 99%

Hypoxia‐induced apoptosis of astrocytes is mediated by reduction of Dicer and activation of caspase‐1

Kang

Kim

Park

et al. 2020

Cell Biology International

View full text Add to dashboard Cite

Hypoxia is a condition in which the whole body or a region of the body is deprived of oxygen supply. The brain is very sensitive to the lack of oxygen and cerebral hypoxia can rapidly cause severe brain damage. Astrocytes are essential for the survival and function of neurons. Therefore, protecting astrocytes against cell death is one of the main therapeutic strategies for treating hypoxia. Hence, the mechanism of hypoxia-induced astrocytic cell death should be fully elucidated. In this study, astrocytes were exposed to hypoxic conditions using a hypoxia work station or the hypoxia mimetic agent cobalt chloride (CoCl 2 ). Both the hypoxic gas mixture (1% O 2 ) and chemical hypoxia-induced apoptotic cell death in T98G glioblastoma cells and mouse primary astrocytes. Reactive oxygen species were generated in response to the hypoxiamediated activation of caspase-1. Active caspase-1 induced the classical caspase-dependent apoptosis of astrocytes. In addition, the microRNA processing enzyme Dicer was cleaved by caspase-3 during hypoxia. Knockdown of Dicer using antisense oligonucleotides induced apoptosis of T98G cells. Taken together, these results suggest that astrocytic cell death during hypoxia is mediated by the reactive oxygen species/caspase-1/classical caspase-dependent apoptotic pathway. In addition, the decrease in Dicer levels by active caspase-3 amplifies this apoptotic pathway via a positive feedback loop. These findings may provide a new target for therapeutic interventions in cerebral hypoxia.

show abstract

Characterization of a naturally occurring truncated Dicer

Cited by 6 publications

References 34 publications

Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer

Nucleotide sequence of miRNA precursor contributes to cleavage site selection by Dicer

Identification of RNA-Binding Proteins as Targetable Putative Oncogenes in Neuroblastoma

Hypoxia‐induced apoptosis of astrocytes is mediated by reduction of Dicer and activation of caspase‐1

Contact Info

Product

Resources

About