Flow-cytometric visualization of C&gt;U mRNA editing reveals the dynamics of the process in live cells

Severi, Francesco; Conticello, Silvestro G.

doi:10.1080/15476286.2015.1026033

Cited by 16 publications

(26 citation statements)

References 75 publications

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“…In order to analyse the function of APOBEC1 in live cells, here we exploit two chimeric APOBEC1 constructs where the position of EGFP, at the N-or the C-terminus, forces the localisation of the chimera to the cytoplasm or the nucleus, respectively (28). These chimeric molecules, coherently with the notion that APOBEC1-mediated RNA editing is a nuclear process, display different RNA-editing activities.…”

Section: Introductionmentioning

confidence: 99%

“…These chimeric molecules, coherently with the notion that APOBEC1-mediated RNA editing is a nuclear process, display different RNA-editing activities. By testing the interplay between wild-type APOBEC1 and these chimeric mutants in live cells using a FACS-based RNA editing assay (28), we find that APOBEC1 dimerisation is dispensable for its activity on RNA, and that proteasomal degradation likely modulates APOBEC1 activity.…”

Section: Introductionmentioning

confidence: 99%

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Dimerisation of APOBEC1 is dispensable for its RNA/DNA editing activity and modulates its availability

Chieca¹,

Montini²,

Severi³

et al. 2018

Preprint

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Among the AID/APOBECs -a family of DNA and RNA deaminases-APOBEC1 physiologically partakes into a complex that edits a CAA codon into UAA Stop codon in the transcript of Apolipoprotein B (ApoB), a protein crucial in the transport of lipids in the blood. Catalytically inactive mutants of APOBEC1 have a dominant negative effect on its activity, as they compete for the targeting to the ApoB mRNA. Here we show that catalytically inactive chimeras of APOBEC1 restricted to different compartments of the cell present different abilities to titrate APOBEC1-mediated RNA editing, and that the ability of APOBEC1 to interact with these mutants is the main determinant for its activity. Our results demonstrate that dimerisation, a feature common to other APOBECs targeting DNA, is not required for APOBEC1 activity on mRNA. Furthermore, APOBEC1-mediated RNA editing is a dynamic process where interplay among the components of the editing complex is regulated through the balance between availability of A1CF, one of APOBEC1 cofactors, and nuclear degradation of APOBEC1.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Dimerisation of APOBEC1 is dispensable for its RNA/DNA editing activity and modulates its availability

Chieca¹,

Montini²,

Severi³

et al. 2018

Preprint

Self Cite

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“…Recombinant A1CF protein was shown to complement APOBEC1 in vitro editing activity and was dependent upon the ApoB mooring sequences (Mehta et al 2000). Over the course of several years, many groups demonstrated the ability of A1CF to complement APOBEC1's editing capacity in vitro, relying on purified recombinant proteins (Mehta and Driscoll 2002;Chester et al 2004) or heterologous expression systems (Chester et al 2003;Sowden et al 2004;Severi and Conticello 2015).…”

Section: Introductionmentioning

confidence: 99%

APOBEC1 complementation factor (A1CF) is dispensable for C-to-U RNA editing in vivo

Snyder

McCarty

Mehalow

et al. 2017

RNA

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Editing of the human and murine ApoB mRNA by APOBEC1, the catalytic enzyme of the protein complex that catalyzes C-to-U RNA editing, creates an internal stop codon within the APOB coding sequence, generating two protein isoforms. It has been long held that APOBEC1-mediated editing activity is dependent on the RNA binding protein A1CF. The function of A1CF in adult tissues has not been reported because a previously reported null allele displays embryonic lethality. This work aimed to address the function of A1CF in adult mouse tissues using a conditional A1cf allele. Unexpectedly, A1cf-null mice were viable and fertile with modest defects in hematopoietic, immune, and metabolic parameters. C-to-U RNA editing was quantified for multiple targets, including ApoB, in the small intestine and liver. In all cases, no changes in RNA editing efficiency were observed. Blood plasma analysis demonstrated a male-specific increase in solute concentration and increased cellularity in the glomeruli of male A1cf-null mice. Urine analysis showed a reduction in solute concentration, suggesting abnormal water homeostasis and possible kidney abnormalities exclusive to the male. Computational identification of kidney C-to-U editing sites from polyadenylated RNA-sequencing identified a number of editing sites exclusive to the kidney. However, molecular analysis of kidney C-to-U editing showed no changes in editing efficiency with A1CF loss. Taken together, these observations demonstrate that A1CF does not act as the APOBEC1 complementation factor in vivo under normal physiological conditions and suggests new roles for A1CF, specifically within the male adult kidney.

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“…with its cofactor catalytic polypeptide-1 (APOBEC1) are required for cellular processes such as the transendothelial migration and the C to U RNA editing [33][34][35][36]. Our study indicates for the first time that miR-25 directly targeted to RBM47.…”

Section: Mir-25 Targets Rna Binding Motif Protein 47mentioning

confidence: 95%

miR-25 Promotes Melanoma Progression by regulating RNA binding motif protein 47

Jiang¹,

Liu²

2018

Med Sci (Paris)

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Melanoma is the most aggressive skin cancer, and accounts for the major part of skin cancer-related deaths in the world. In addition, the underlying mechanism of tumor progression in melanoma remains far from being elucidated. In this study, we have evaluated the function of miR-25 in melanoma. First, we examined the expression of miR-25 in four melanoma cell lines (A875, MV3, M14 and uacc-257) and in a normal melanocyte cell line (HEM-a). Then, we overexpressed miR-25 in M14 cells. Our results show that miR-25 promotes M14 cell proliferation and migration. We found that miR-25 up-regulates the PI3K/Akt/mTOR signaling pathway in these tumor cells. Furthermore, a luciferase-based reporter gene assay showed that miR-25 could directly target the RNA-binding motif protein 47 (RBM47). Taken together, our findings suggest that RBM47 is a promising target for the treatment of melanoma.

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Flow-cytometric visualization of C>U mRNA editing reveals the dynamics of the process in live cells

Cited by 16 publications

References 75 publications

Dimerisation of APOBEC1 is dispensable for its RNA/DNA editing activity and modulates its availability

Dimerisation of APOBEC1 is dispensable for its RNA/DNA editing activity and modulates its availability

APOBEC1 complementation factor (A1CF) is dispensable for C-to-U RNA editing in vivo

miR-25 Promotes Melanoma Progression by regulating RNA binding motif protein 47

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