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Alternative Splicing at a NAGNAG Acceptor Site as a Novel Phenotype Modifier


Approximately 30% of alleles causing genetic disorders generate premature termination codons (PTCs), which are usually associated with severe phenotypes. However, bypassing the deleterious stop codon can lead to a mild disease outcome. Splicing at NAGNAG tandem splice sites has been reported to result in insertion or deletion (indel) of three nucleotides. We identified such a mechanism as the origin of the mild to asymptomatic phenotype observed in cystic fibrosis patients homozygous for the E831X mutation (2623G>T) in the CFTR gene. Analyses performed on nasal epithelial cell mRNA detected three distinct isoforms, a considerably more complex situation than expected for a single nucleotide substitution. Structure-function studies and in silico analyses provided the first experimental evidence of an indel of a stop codon by alternative splicing at a NAGNAG acceptor site. In addition to contributing to proteome plasticity, alternative splicing at a NAGNAG tandem site can thus remove a disease-causing UAG stop codon. This molecular study reveals a naturally occurring mechanism where the effect of either modifier genes or epigenetic factors could be suspected. This finding is of importance for genetic counseling as well as for deciding appropriate therapeutic strategies.


Vyšlo v časopise: Alternative Splicing at a NAGNAG Acceptor Site as a Novel Phenotype Modifier. PLoS Genet 6(10): e32767. doi:10.1371/journal.pgen.1001153
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1001153

Souhrn

Approximately 30% of alleles causing genetic disorders generate premature termination codons (PTCs), which are usually associated with severe phenotypes. However, bypassing the deleterious stop codon can lead to a mild disease outcome. Splicing at NAGNAG tandem splice sites has been reported to result in insertion or deletion (indel) of three nucleotides. We identified such a mechanism as the origin of the mild to asymptomatic phenotype observed in cystic fibrosis patients homozygous for the E831X mutation (2623G>T) in the CFTR gene. Analyses performed on nasal epithelial cell mRNA detected three distinct isoforms, a considerably more complex situation than expected for a single nucleotide substitution. Structure-function studies and in silico analyses provided the first experimental evidence of an indel of a stop codon by alternative splicing at a NAGNAG acceptor site. In addition to contributing to proteome plasticity, alternative splicing at a NAGNAG tandem site can thus remove a disease-causing UAG stop codon. This molecular study reveals a naturally occurring mechanism where the effect of either modifier genes or epigenetic factors could be suspected. This finding is of importance for genetic counseling as well as for deciding appropriate therapeutic strategies.


Zdroje

1. HolbrookJA

Neu-YilikG

HentzeMW

KulozikAE

2004

Nonsense-mediated decay approaches the clinic.

Nat Genet

36

801

808

2. MaserRS

ZinkelR

PetriniJH

2001

An alternative mode of translation permits production of a variant NBS1 protein from the common Nijmegen breakage syndrome allele.

Nat Genet

27

417

421

3. MaquatLE

1995

When cells stop making sense: effects of nonsense codons on RNA metabolism in vertebrate cells.

RNA

1

453

465

4. ValentineCR

1998

The association of nonsense codons with exon skipping.

Mutat Res

411

87

117

5. HillerM

HuseK

SzafranskiK

JahnN

HampeJ

2004

Widespread occurrence of alternative splicing at NAGNAG acceptors contributes to proteome plasticity.

Nat Genet

36

1255

1257

6. SugnetCW

KentWJ

AresMJr

HausslerD

2004

Transcriptome and genome conservation of alternative splicing events in humans and mice.

Pac Symp Biocomput

66

77

7. ZavolanM

KondoS

SchonbachC

AdachiJ

HumeDA

2003

Impact of alternative initiation, splicing, and termination on the diversity of the mRNA transcripts encoded by the mouse transcriptome.

Genome Res

13

1290

1300

8. MaugeriA

van DrielMA

van de PolDJ

KleveringBJ

van HarenFJ

1999

The 2588G→C mutation in the ABCR gene is a mild frequent founder mutation in the Western European population and allows the classification of ABCR mutations in patients with Stargardt disease.

Am J Hum Genet

64

1024

1035

9. HillerM

PlatzerM

2008

Widespread and subtle: alternative splicing at short-distance tandem sites.

Trends Genet

24

246

255

10. RiordanJR

RommensJM

KeremB

AlonN

RozmahelR

1989

Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA.

Science

245

1066

1073

11. Nissim-RafiniaM

KeremB

2002

Splicing regulation as a potential genetic modifier.

Trends Genet

18

123

127

12. DupuitF

KalinN

BrezillonS

HinnraskyJ

TummlerB

1995

CFTR and differentiation markers expression in non-CF and delta F 508 homozygous CF nasal epithelium.

J Clin Invest

96

1601

1611

13. LindeL

BoelzS

Nissim-RafiniaM

OrenYS

WilschanskiM

2007

Nonsense-mediated mRNA decay affects nonsense transcript levels and governs response of cystic fibrosis patients to gentamicin.

J Clin Invest

117

683

692

14. HullJ

ShackletonS

HarrisA

1994

Analysis of mutations and alternative splicing patterns in the CFTR gene using mRNA derived from nasal epithelial cells.

Hum Mol Genet

3

1141

1146

15. ChengSH

GregoryRJ

MarshallJ

PaulS

SouzaDW

1990

Defective intracellular transport and processing of CFTR is the molecular basis of most cystic fibrosis.

Cell

63

827

834

16. VerlingueC

MercierB

LecoqI

AudrezetMP

LarocheD

1994

Retrospective study of the cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations in Guthrie cards from a large cohort of neonatal screening for cystic fibrosis.

Hum Genet

93

429

434

17. VaronR

MagdorfK

StaabD

WahnHU

KrawczakM

1995

Recurrent nasal polyps as a monosymptomatic form of cystic fibrosis associated with a novel in-frame deletion (591del18) in the CFTR gene.

Hum Mol Genet

4

1463

1464

18. DayangacD

ErdemH

YilmazE

SahinA

SohnC

2004

Mutations of the CFTR gene in Turkish patients with congenital bilateral absence of the vas deferens.

Hum Reprod

19

1094

1100

19. Di GirgentiC

VirrusoL

MessineoR

CannuscioA

TerminiL

2008

From “evocative” symptoms to genotype deltaF508/E831X.

Journal of Cystic Fibrosis

7

S11

S11

20. GrangeiaA

SaR

CarvalhoF

MartinJ

GirodonE

2007

Molecular characterization of the cystic fibrosis transmembrane conductance regulator gene in congenital absence of the vas deferens.

Genet Med

9

163

172

21. Rave-HarelN

KeremE

Nissim-RafiniaM

MadjarI

GoshenR

1997

The molecular basis of partial penetrance of splicing mutations in cystic fibrosis.

Am J Hum Genet

60

87

94

22. NoonePG

PueCA

ZhouZ

FriedmanKJ

WakelingEL

2000

Lung disease associated with the IVS8 5T allele of the CFTR gene.

Am J Respir Crit Care Med

162

1919

1924

23. RamalhoAS

BeckS

MeyerM

PenqueD

CuttingGR

2002

Five percent of normal cystic fibrosis transmembrane conductance regulator mRNA ameliorates the severity of pulmonary disease in cystic fibrosis.

Am J Respir Cell Mol Biol

27

619

627

24. TsaiKW

TarnWY

LinWC

2007

Wobble splicing reveals the role of the branch point sequence-to-NAGNAG region in 3′ tandem splice site selection.

Mol Cell Biol

27

5835

5848

25. AkermanM

Mandel-GutfreundY

2006

Alternative splicing regulation at tandem 3′ splice sites.

Nucleic Acids Res

34

23

31

26. HillerM

HuseK

SzafranskiK

JahnN

HampeJ

2006

Single-nucleotide polymorphisms in NAGNAG acceptors are highly predictive for variations of alternative splicing.

Am J Hum Genet

78

291

302

27. SinhaR

NikolajewaS

SzafranskiK

HillerM

JahnN

2009

Accurate prediction of NAGNAG alternative splicing.

Nucleic Acids Res

37

3569

3579

28. HillerM

SzafranskiK

BackofenR

PlatzerM

2006

Alternative splicing at NAGNAG acceptors: simply noise or noise and more?

PLoS Genet

2

e207

doi:10.1371/journal.pgen.0020207

29. LivakKJ

SchmittgenTD

2001

Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

Methods

25

402

408

30. FanenP

ClainJ

LabartheR

HulinP

GirodonE

1999

Structure-function analysis of a double-mutant cystic fibrosis transmembrane conductance regulator protein occurring in disorders related to cystic fibrosis.

FEBS Lett

452

371

374

31. TanguyG

DrevillonL

ArousN

HasnainA

HinzpeterA

2008

CSN5 binds to misfolded CFTR and promotes its degradation.

Biochim Biophys Acta

1783

1189

1199

32. CaciE

CaputoA

HinzpeterA

ArousN

FanenP

2008

Evidence for direct CFTR inhibition by CFTR(inh)-172 based on Arg347 mutagenesis.

Biochem J

413

135

142

33. GaliettaLJ

HaggiePM

VerkmanAS

2001

Green fluorescent protein-based halide indicators with improved chloride and iodide affinities.

FEBS Lett

499

220

224

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Genetika Reprodukčná medicína

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PLOS Genetics


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Autori: MUDr. Tomáš Ürge, PhD.

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