Long CTG Tracts from the Myotonic Dystrophy Gene Induce Deletions and Rearrangements during Recombination at the APRT Locus in CHO Cells
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| Published in: | Molecular and Cellular Biology Vol. 23; no. 9; pp. 3152 - 3162 |
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| AbstractList | Expansion of CTG triplet repeats in the 3' untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG repeats is thought to arise from their capacity to form hairpin DNA structures. How these structures interact with various aspects of DNA metabolism has been studied intensely for Escherichia coli and Saccharomyces cerevisiae but is relatively uncharacterized in mammalian cells. To examine the stability of (CTG) sub(17), (CTG) sub(98), and (CTG) sub(183) repeats during homologous recombination, we placed them in the second intron of one copy of a tandemly duplicated pair of APRT genes. Cells selected for homologous recombination between the two copies of the APRT gene displayed distinctive patterns of change. Among recombinants from cells with (CTG) sub(98) and (CTG) sub(183), 5% had lost large numbers of repeats and 10% had suffered rearrangements, a frequency more than 50-fold above normal levels. Analysis of individual rearrangements confirmed the involvement of the CTG repeats. Similar changes were not observed in proliferating (CTG) sub(98) and (CTG) sub(183) cells that were not recombinant at APRT. Instead, they displayed high frequencies of small changes in repeat number. The (CTG) sub(17) repeats were stable in all assays. These studies indicate that homologous recombination strongly destabilizes long tracts of CTG repeats. Expansion of CTG triplet repeats in the 3′ untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG repeats is thought to arise from their capacity to form hairpin DNA structures. How these structures interact with various aspects of DNA metabolism has been studied intensely for Escherichia coli and Saccharomyces cerevisiae but is relatively uncharacterized in mammalian cells. To examine the stability of (CTG) 17 , (CTG) 98 , and (CTG) 183 repeats during homologous recombination, we placed them in the second intron of one copy of a tandemly duplicated pair of APRT genes. Cells selected for homologous recombination between the two copies of the APRT gene displayed distinctive patterns of change. Among recombinants from cells with (CTG) 98 and (CTG) 183 , 5% had lost large numbers of repeats and 10% had suffered rearrangements, a frequency more than 50-fold above normal levels. Analysis of individual rearrangements confirmed the involvement of the CTG repeats. Similar changes were not observed in proliferating (CTG) 98 and (CTG) 183 cells that were not recombinant at APRT . Instead, they displayed high frequencies of small changes in repeat number. The (CTG) 17 repeats were stable in all assays. These studies indicate that homologous recombination strongly destabilizes long tracts of CTG repeats. Expansion of CTG triplet repeats in the 3′ untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG repeats is thought to arise from their capacity to form hairpin DNA structures. How these structures interact with various aspects of DNA metabolism has been studied intensely for Escherichia coli and Saccharomyces cerevisiae but is relatively uncharacterized in mammalian cells. To examine the stability of (CTG) 17 , (CTG) 98 , and (CTG) 183 repeats during homologous recombination, we placed them in the second intron of one copy of a tandemly duplicated pair of APRT genes. Cells selected for homologous recombination between the two copies of the APRT gene displayed distinctive patterns of change. Among recombinants from cells with (CTG) 98 and (CTG) 183 , 5% had lost large numbers of repeats and 10% had suffered rearrangements, a frequency more than 50-fold above normal levels. Analysis of individual rearrangements confirmed the involvement of the CTG repeats. Similar changes were not observed in proliferating (CTG) 98 and (CTG) 183 cells that were not recombinant at APRT. Instead, they displayed high frequencies of small changes in repeat number. The (CTG) 17 repeats were stable in all assays. These studies indicate that homologous recombination strongly destabilizes long tracts of CTG repeats. Article Usage Stats Services MCB Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue MCB About MCB Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy MCB RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0270-7306 Online ISSN: 1098-5549 Copyright © 2014 by the American Society for Microbiology. For an alternate route to MCB .asm.org, visit: MCB Expansion of CTG triplet repeats in the 3' untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG repeats is thought to arise from their capacity to form hairpin DNA structures. How these structures interact with various aspects of DNA metabolism has been studied intensely for Escherichia coli and Saccharomyces cerevisiae but is relatively uncharacterized in mammalian cells. To examine the stability of (CTG)(17), (CTG)(98), and (CTG)(183) repeats during homologous recombination, we placed them in the second intron of one copy of a tandemly duplicated pair of APRT genes. Cells selected for homologous recombination between the two copies of the APRT gene displayed distinctive patterns of change. Among recombinants from cells with (CTG)(98) and (CTG)(183), 5% had lost large numbers of repeats and 10% had suffered rearrangements, a frequency more than 50-fold above normal levels. Analysis of individual rearrangements confirmed the involvement of the CTG repeats. Similar changes were not observed in proliferating (CTG)(98) and (CTG)(183) cells that were not recombinant at APRT. Instead, they displayed high frequencies of small changes in repeat number. The (CTG)(17) repeats were stable in all assays. These studies indicate that homologous recombination strongly destabilizes long tracts of CTG repeats. |
| Author | Ravi R. Iyer R. Geoffrey Sargent James L. Meservy Gregory J. McKenzie Fung Chan John H. Wilson Robert D. Wells |
| AuthorAffiliation | Verna and Marrs McLean Department of Biochemistry and Molecular Biology, 1 Department of Molecular and Human Genetics, Baylor College of Medicine, 3 Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University, Houston, Texas 77030 2 |
| AuthorAffiliation_xml | – name: Verna and Marrs McLean Department of Biochemistry and Molecular Biology, 1 Department of Molecular and Human Genetics, Baylor College of Medicine, 3 Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University, Houston, Texas 77030 2 |
| Author_xml | – sequence: 1 givenname: James L. surname: Meservy fullname: Meservy, James L. organization: Verna and Marrs McLean Department of Biochemistry and Molecular Biology – sequence: 2 givenname: R. Geoffrey surname: Sargent fullname: Sargent, R. Geoffrey organization: Verna and Marrs McLean Department of Biochemistry and Molecular Biology – sequence: 3 givenname: Ravi R. surname: Iyer fullname: Iyer, Ravi R. organization: Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University – sequence: 4 givenname: Fung surname: Chan fullname: Chan, Fung organization: Verna and Marrs McLean Department of Biochemistry and Molecular Biology – sequence: 5 givenname: Gregory J. surname: McKenzie fullname: McKenzie, Gregory J. organization: Department of Molecular and Human Genetics, Baylor College of Medicine – sequence: 6 givenname: Robert D. surname: Wells fullname: Wells, Robert D. organization: Institute of Biosciences and Technology, Center for Genome Research, Texas A & M University – sequence: 7 givenname: John H. surname: Wilson fullname: Wilson, John H. email: jwilson@bcm.tmc.edu organization: Department of Molecular and Human Genetics, Baylor College of Medicine |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/12697816$$D View this record in MEDLINE/PubMed |
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| Notes | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 Corresponding author. Mailing address: Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030. Phone: (713) 798-5760. Fax: (713) 796-9438. E-mail: jwilson@bcm.tmc.edu. |
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Mendeley... Expansion of CTG triplet repeats in the 3′ untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG... Expansion of CTG triplet repeats in the 3' untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG... Expansion of CTG triplet repeats in the 3′ untranslated region of the DMPK gene causes the autosomal dominant disorder myotonic dystrophy. Instability of CTG... |
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| StartPage | 3152 |
| SubjectTerms | Adenine Phosphoribosyltransferase - genetics Animals Cell Line CHO Cells Cricetinae DNA Dynamics and Chromosome Structure Gene Dosage Gene Rearrangement Humans Introns Myotonin-Protein Kinase Protein-Serine-Threonine Kinases - genetics Recombination, Genetic Sequence Deletion Trinucleotide Repeat Expansion |
| Title | Long CTG Tracts from the Myotonic Dystrophy Gene Induce Deletions and Rearrangements during Recombination at the APRT Locus in CHO Cells |
| URI | http://mcb.asm.org/content/23/9/3152.abstract https://www.tandfonline.com/doi/abs/10.1128/MCB.23.9.3152-3162.2003 https://www.ncbi.nlm.nih.gov/pubmed/12697816 https://search.proquest.com/docview/18733900 https://pubmed.ncbi.nlm.nih.gov/PMC153196 |
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