Introducing sense into nonsense in treatments of human genetic diseases

Linde, Liat; Kerem, Batsheva

doi:10.1016/j.tig.2008.08.010

Cited by 186 publications

(189 citation statements)

References 104 publications

Supporting

Mentioning

179

Contrasting

Unclassified

Order By: Relevance

“…Read-through therapies have shown promise in generating full-length proteins without concomitantly stabilizing PTC-bearing mRNAs (26,27). However, a deeper understanding of how such transcripts are recognized and selectively degraded by NMD is needed considering that UPF1 has been reported to bind to many mammalian mRNAs regardless of PTC status (12,28).…”

mentioning

confidence: 99%

Rules that govern UPF1 binding to mRNA 3′ UTRs

Kurosaki

Maquat

2013

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

112

120

View full text Add to dashboard Cite

Nonsense-mediated mRNA decay (NMD), which degrades transcripts harboring a premature termination codon (PTC), depends on the helicase up-frameshift 1 (UPF1). However, mRNAs that are not NMD targets also bind UPF1. What governs the timing, position, and function of UPF1 binding to mRNAs remains unclear. We provide evidence that (i) multiple UPF1 molecules accumulate on the 3′-untranslated region (3′ UTR) of PTC-containing mRNAs and to an extent that is greater per unit 3′ UTR length if the mRNA is an NMD target; (ii) UPF1 binding begins ≥35 nt downstream of the PTC; (iii) enhanced UPF1 binding to the 3′ UTR of PTC-containing mRNA relative to its PTC-free counterpart depends on translation; and (iv) the presence of a 3′ UTR exon-junction complex (EJC) further enhances UPF1 binding and/or affinity. Our data suggest that NMD involves UPF1 binding along a 3′ UTR whether the 3′ UTR contains an EJC. This binding explains how mRNAs without a 3′ UTR EJC but with an abnormally long 3′ UTR can be NMD targets, albeit not as efficiently as their counterparts that contain a 3′ UTR EJC.messenger ribonucleoprotein structure | RNA-binding protein | messenger RNA quality control I n mammalian cells, mRNAs and the pre-mRNAs from which they derive continually lose and acquire proteins in ways that reflect past and current metabolic states and influence future metabolic steps. For example, newly synthesized mRNAs, which support the pioneer round of translation, are bound by the capbinding protein (CBP) heterodimer CBP80−CBP20 and, provided they underwent splicing, exon-junction complexes (EJCs) (1). In contrast, the bulk of cellular mRNAs have lost CBP80−CBP20 and EJCs, have acquired eukaryotic translation initiation factor (eIF)4E in the place of CBP80−CBP20, and support most of cellular translation. CBP80 and EJCs are important for nonsensemediated mRNA decay (NMD), which down-regulates mRNAs that harbor either an exon-exon junction downstream of a termination codon (2, 3) and/or an abnormally long 3′ UTR (3-6). Thus, NMD targets newly synthesized mRNAs during the pioneer round of translation, whereas eIF4E-bound mRNAs appear to be immune to NMD (1, 3). This conclusion is supported by single-RNA fluorescent in situ-hybridization measurements of premature termination codon (PTC)-containing mRNAs in intact cells after the induction of mRNA synthesis (7).Messenger ribonucleoprotein (mRNP) remodeling also takes place during NMD. When translation terminates at a PTC and an EJC is situated sufficiently downstream of the PTC so as not to be displaced by the terminating ribosome (8, 9), a complex called suppressor with morphogenic effect on genitalia (SMG) 1−upframeshift 1 (UPF1)−eukaryotic release factor (eRF) 1−eRF3 (SURF) is thought to form at the PTC and, together with the downstream EJC, constitutes a decay-inducing complex (DECID) (10, 11). The transient and/or weak interaction of mRNA capbound CBP80 with UPF1, which occurs on mRNAs even if they are not NMD targets, promotes UPF1 binding to eRF1−eRF3 and, subsequently, to an EJC (12...

show abstract

mentioning

confidence: 99%

Rules that govern UPF1 binding to mRNA 3′ UTRs

Kurosaki

Maquat

2013

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

112

120

View full text Add to dashboard Cite

show abstract

“…1). Since one of ) the alleles encodes a premature stop codon [8], we had expected a decline in overall RPI mRNA due to activity of the nonsense mediated decay [18]. However, all qPCR experiments indicated a reduction of the RPI mRNA below the 50% margin.…”

Section: Resultsmentioning

confidence: 97%

The difference between rare and exceptionally rare: molecular characterization of ribose 5-phosphate isomerase deficiency

et al. 2010

View full text Add to dashboard Cite

Ribose 5-phosphate isomerase (RPI) deficiency is an enzymopathy of the pentose phosphate pathway. It manifests with progressive leukoencephalopathy and peripheral neuropathy and belongs, with one sole diagnosed case, to the rarest human disorders. The single patient was found compound heterozygous for a RPI frameshift and a missense (RPI Ala61Val ) allele. Here, we report that two patient-derived cell lines differ in RPI enzyme activity, enzyme concentration, and mRNA expression. Furthermore, we present a transgenic yeast model, which exhibits metabolite-and enzyme-activity changes that correspond to the human syndrome and show that the decrease in RPI activity in patient cells is not fully attributable to the residue exchange. Taken together, our results demonstrate that RPI deficiency is caused by the combination of a RPI null allele with an allele that encodes for a partially active enzyme which has, in addition, cell-type-dependent expression deficits. We speculate that a low probability for comparable traits accounts for the rareness of RPI deficiency.

show abstract

“…This is similar to results obtained with aminoglycosides and PTC124 (drugs tested in clinical trials) in cultured cell lines and mouse models of several diseases, where the efficiency of production of full-length functional protein ranged from 1 to 25%. 15,16 Reasons for this low efficiency are the dependence on PTC identity, 39 PTC context, 40 activity of translation release factor 1 (eRF1) 21 and the decoding efficiency of the sup-tRNA. 40 A question raised upon use of readthrough strategies is its effect on global translation efficiency, namely the possibility of suppressing canonical stop codons.…”

Section: Resultsmentioning

confidence: 99%

Rescue of wild-type E-cadherin expression from nonsense-mutated cancer cells by a suppressor-tRNA

et al. 2014

View full text Add to dashboard Cite

Hereditary diffuse gastric cancer (HDGC) syndrome, although rare, is highly penetrant at an early age, and is severe and incurable because of ineffective screening tools and therapy. Approximately 45% of HDGC families carry germline CDH1/Ecadherin alterations, 20% of which are nonsense leading to premature protein truncation. Prophylactic gastrectomy is the only recommended approach for all asymptomatic CDH1 mutation carriers. Suppressor-tRNAs can replace premature stop codons (PTCs) with a cognate amino acid, inducing readthrough and generating full-length proteins. The use of suppressor-tRNAs in HDGC patients could therefore constitute a less invasive therapeutic option for nonsense mutation carriers, delaying the development of gastric cancer. Our analysis revealed that 23/108 (21.3%) of E-cadherin-mutant families carried nonsense mutations that could be potentially corrected by eight suppressor-tRNAs, and arginine was the most frequently affected amino acid. Using site-directed mutagenesis, we developed an arginine suppressor-tRNA vector to correct one HDGC nonsense mutation. E-cadherin-deficient cell lines were transfected with plasmids carrying simultaneously the suppressor-tRNA and wild-type or mutant CDH1 mini-genes. RT-PCR, western blot, immunofluorescence, flow cytometry and proximity ligation assay (PLA) were used to establish the model, and monitor mRNA and protein expression and function recovery from CDH1 vectors. Cells expressing a CDH1 mini-gene, carrying a nonsense mutation and the suppressor-tRNA, recovered full-length E-cadherin expression and its correct localization and incorporation into the adhesion complex. This is the first demonstration of functional recovery of a mutated causative gene in hereditary cancer by cognate amino acid replacement with suppressor-tRNAs. Of the HDGC families, 21.3% are candidates for correction with suppressor-tRNAs to potentially delay cancer onset.

show abstract

Introducing sense into nonsense in treatments of human genetic diseases

Cited by 186 publications

References 104 publications

Rules that govern UPF1 binding to mRNA 3′ UTRs

Rules that govern UPF1 binding to mRNA 3′ UTRs

The difference between rare and exceptionally rare: molecular characterization of ribose 5-phosphate isomerase deficiency

Rescue of wild-type E-cadherin expression from nonsense-mutated cancer cells by a suppressor-tRNA

Contact Info

Product

Resources

About