Counting CAG Repeats in the Huntington's Disease Gene by Restriction Endonuclease EcoP15I Cleavage

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2002 Aug 15

PMID 12177311

Citations 18

Authors

Elisabeth Moncke-Buchner

Stefanie Reich

Merlind Mucke

Monika Reuter

Walter Messer

Erich E Wanker

Detlev H Kruger

Affiliations

Soon will be listed here.

Abstract

Huntington's disease (HD) is a progressive neurodegenerative disorder with autosomal-dominant inheritance. The disease is caused by a CAG trinucleotide repeat expansion located in the first exon of the HD gene. The CAG repeat is highly polymorphic and varies from 6 to 37 repeats on chromosomes of unaffected individuals and from more than 30 to 180 repeats on chromosomes of HD patients. In this study, we show that the number of CAG repeats in the HD gene can be determined by restriction of the DNA with the endonuclease EcoP15I and subsequent analysis of the restriction fragment pattern by electrophoresis through non-denaturing polyacrylamide gels using the ALFexpress DNA Analysis System. CAG repeat numbers in the normal (30 and 35 repeats) as well as in the pathological range (81 repeats) could be accurately counted using this assay. Our results suggest that this high-resolution method can be used for the exact length determination of CAG repeats in HD genes as well as in genes affected in related CAG repeat disorders.

Citing Articles

Genetic landscape of congenital pouch colon: systematic review and functional enrichment study.

Phugat S, Sharma J, Kumar S, Jain V, Dhua A, Yadav D Pediatr Surg Int. 2024; 40(1):314.

PMID: 39557707 DOI: 10.1007/s00383-024-05878-8.

Therapeutic approaches targeting aging and cellular senescence in Huntington's disease.

Bhat A, Moglad E, Afzal M, Thapa R, Almalki W, Kazmi I CNS Neurosci Ther. 2024; 30(10):e70053.

PMID: 39428700 PMC: 11491556. DOI: 10.1111/cns.70053.

Generation of Rhesus Macaque Embryos with Expanded CAG Trinucleotide Repeats in the Gene.

Ryu J, Statz J, Chan W, Oyama K, Custer M, Wienisch M Cells. 2024; 13(10.

PMID: 38786052 PMC: 11119628. DOI: 10.3390/cells13100829.

Replication Protein A, the Main Eukaryotic Single-Stranded DNA Binding Protein, a Focal Point in Cellular DNA Metabolism.

Nasheuer H, Meaney A, Hulshoff T, Thiele I, Onwubiko N Int J Mol Sci. 2024; 25(1).

PMID: 38203759 PMC: 10779431. DOI: 10.3390/ijms25010588.

Benchmarking long-read genome sequence alignment tools for human genomics applications.

LoTempio J, Delot E, Vilain E PeerJ. 2023; 11:e16515.

PMID: 38130927 PMC: 10734412. DOI: 10.7717/peerj.16515.

References

Matsuura T, Yamagata T, Burgess D, Rasmussen A, Grewal R, Watase K . Large expansion of the ATTCT pentanucleotide repeat in spinocerebellar ataxia type 10. Nat Genet. 2000; 26(2):191-4. DOI: 10.1038/79911. View

Wanker E . Protein aggregation and pathogenesis of Huntington's disease: mechanisms and correlations. Biol Chem. 2000; 381(9-10):937-42. DOI: 10.1515/BC.2000.114. View

Tobin A, Signer E . Huntington's disease: the challenge for cell biologists. Trends Cell Biol. 2000; 10(12):531-6. DOI: 10.1016/s0962-8924(00)01853-5. View

Roberts R, Macelis D . REBASE--restriction enzymes and methylases. Nucleic Acids Res. 2000; 29(1):268-9. PMC: 29853. DOI: 10.1093/nar/29.1.268. View

van Belkum A . Short sequence repeats in microbial pathogenesis and evolution. Cell Mol Life Sci. 2001; 56(9-10):729-34. PMC: 11146772. DOI: 10.1007/s000180050019. View

Gusella J, Macdonald M . Molecular genetics: unmasking polyglutamine triggers in neurodegenerative disease. Nat Rev Neurosci. 2001; 1(2):109-15. DOI: 10.1038/35039051. View

Waelter S, Boeddrich A, Lurz R, Scherzinger E, Lueder G, Lehrach H . Accumulation of mutant huntingtin fragments in aggresome-like inclusion bodies as a result of insufficient protein degradation. Mol Biol Cell. 2001; 12(5):1393-407. PMC: 34592. DOI: 10.1091/mbc.12.5.1393. View

Pelotti S, Maiolini E, Bini C, Rimondi S, Luiselli D, Pappalardo G . Automated fluorescence analysis of CAG repeats at the human androgen receptor gene (HUMARA): evaluation of polymorphism in an Italian sample and report of a new allele. Am J Forensic Med Pathol. 2001; 22(1):55-7. DOI: 10.1097/00000433-200103000-00010. View

Dryden D, Murray N, Rao D . Nucleoside triphosphate-dependent restriction enzymes. Nucleic Acids Res. 2001; 29(18):3728-41. PMC: 55918. DOI: 10.1093/nar/29.18.3728. View

10.

Mucke M, Reich S, Reuter M, Kruger D . DNA cleavage by type III restriction-modification enzyme EcoP15I is independent of spacer distance between two head to head oriented recognition sites. J Mol Biol. 2001; 312(4):687-98. DOI: 10.1006/jmbi.2001.4998. View

11.

Wallerand H, Remy-Martin A, Chabannes E, Bermont L, Adessi G, Bittard H . Relationship between expansion of the CAG repeat in exon 1 of the androgen receptor gene and idiopathic male infertility. Fertil Steril. 2001; 76(4):769-74. DOI: 10.1016/s0015-0282(01)01987-2. View

12.

Choudhry S, Mukerji M, Srivastava A, Jain S, Brahmachari S . CAG repeat instability at SCA2 locus: anchoring CAA interruptions and linked single nucleotide polymorphisms. Hum Mol Genet. 2001; 10(21):2437-46. DOI: 10.1093/hmg/10.21.2437. View

13.

Ishii T, Sato S, Kosaki K, Sasaki G, Muroya K, Ogata T . Micropenis and the AR Gene: mutation and CAG repeat-length analysis. J Clin Endocrinol Metab. 2001; 86(11):5372-8. DOI: 10.1210/jcem.86.11.7999. View

14.

Bickle T, Kruger D . Biology of DNA restriction. Microbiol Rev. 1993; 57(2):434-50. PMC: 372918. DOI: 10.1128/mr.57.2.434-450.1993. View

15.

Warner J, Barron L, Brock D . A new polymerase chain reaction (PCR) assay for the trinucleotide repeat that is unstable and expanded on Huntington's disease chromosomes. Mol Cell Probes. 1993; 7(3):235-9. DOI: 10.1006/mcpr.1993.1034. View

16.

Duyao M, Ambrose C, Myers R, Novelletto A, Persichetti F, Frontali M . Trinucleotide repeat length instability and age of onset in Huntington's disease. Nat Genet. 1993; 4(4):387-92. DOI: 10.1038/ng0893-387. View

17.

Snell R, MacMillan J, Cheadle J, Fenton I, Lazarou L, Davies P . Relationship between trinucleotide repeat expansion and phenotypic variation in Huntington's disease. Nat Genet. 1993; 4(4):393-7. DOI: 10.1038/ng0893-393. View

18.

Andrew S, Goldberg Y, Kremer B, Telenius H, Theilmann J, Adam S . The relationship between trinucleotide (CAG) repeat length and clinical features of Huntington's disease. Nat Genet. 1993; 4(4):398-403. DOI: 10.1038/ng0893-398. View

19.

Meisel A, Mackeldanz P, Bickle T, Kruger D, Schroeder C . Type III restriction endonucleases translocate DNA in a reaction driven by recognition site-specific ATP hydrolysis. EMBO J. 1995; 14(12):2958-66. PMC: 398416. DOI: 10.1002/j.1460-2075.1995.tb07296.x. View

20.

Kruger D, Kupper D, Meisel A, Reuter M, Schroeder C . The significance of distance and orientation of restriction endonuclease recognition sites in viral DNA genomes. FEMS Microbiol Rev. 1995; 17(1-2):177-84. DOI: 10.1111/j.1574-6976.1995.tb00200.x. View