Single base discrimination of CENP-B repeats on mouse and human Chromosomes with PNA-FISH.
Identifieur interne : 003783 ( Main/Exploration ); précédent : 003782; suivant : 003784Single base discrimination of CENP-B repeats on mouse and human Chromosomes with PNA-FISH.
Auteurs : C. Chen [États-Unis] ; Y K Hong ; S D Ontiveros ; M. Egholm ; W M StraussSource :
- Mammalian genome : official journal of the International Mammalian Genome Society [ 0938-8990 ] ; 1999.
Descripteurs français
- KwdFr :
- Acides nucléiques peptidiques, Amorces ADN, Animaux, Appariement de bases, Autoantigènes, Humains, Protéine B du centromère, Protéines chromosomiques nonhistones (génétique), Protéines de liaison à l'ADN, Sensibilité et spécificité, Souris, Souris de lignée C57BL, Spécificité d'espèce, Séquences répétées d'acides nucléiques, Technique FISH ().
- MESH :
- génétique : Protéines chromosomiques nonhistones.
- Acides nucléiques peptidiques, Amorces ADN, Animaux, Appariement de bases, Autoantigènes, Humains, Protéine B du centromère, Protéines de liaison à l'ADN, Sensibilité et spécificité, Souris, Souris de lignée C57BL, Spécificité d'espèce, Séquences répétées d'acides nucléiques, Technique FISH.
English descriptors
- KwdEn :
- Animals, Autoantigens, Base Pairing, Centromere Protein B, Chromosomal Proteins, Non-Histone (genetics), DNA Primers, DNA-Binding Proteins, Humans, In Situ Hybridization, Fluorescence (methods), Mice, Mice, Inbred C57BL, Peptide Nucleic Acids, Repetitive Sequences, Nucleic Acid, Sensitivity and Specificity, Species Specificity.
- MESH :
- chemical , genetics : Chromosomal Proteins, Non-Histone.
- chemical : Autoantigens, Centromere Protein B, DNA Primers, DNA-Binding Proteins, Peptide Nucleic Acids.
- methods : In Situ Hybridization, Fluorescence.
- Animals, Base Pairing, Humans, Mice, Mice, Inbred C57BL, Repetitive Sequences, Nucleic Acid, Sensitivity and Specificity, Species Specificity.
Abstract
The satellite repeat structure of the mammalian centromere contains the CENP-B protein binding site. Using the peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH), we show by direct PNA-DNA binding that all detectable CENP-B sites in a mammalian genome might have the same sequence. Two species-specific PNA 17-mers, pMm and pMc, were identified from CENP-B binding sites of Mus musculus and M. caroli, respectively. Fluorescence in situ hybridization confirmed that pMc hybridized to M. caroli centromeres only; however, pMm cross-hybridized to M. musculus and human centromeres. By using a series of CENP-B PNA 17-mers containing 1, 2, 3, 5, and 7 base-pair mismatches to their DNA counterparts, we further demonstrate that PNA-FISH can discriminate between two CENP-B DNA sequences that differ by a single base-pair in mouse and human centromeres, suggesting the degree of conservation of CENP-B sequences throughout the genome. In comparison with DNA oligonucleotides, PNA oligomers demonstrate the higher sequence specificity, improved stability, reproducibility, and lower background. Therefore, PNA oligomers have significant advantages over DNA oligonucleotide probes in analyzing microsatellites in a genome.
DOI: 10.1007/s003359900934
PubMed: 9892726
Affiliations:
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Chen</name><affiliation wicri:level="1"><nlm:affiliation>Harvard Institutes of Human Genetics, Harvard Medical School, Beth Israel Deaconess Medical Center, 4 Blackfan Circle, Boston, Massachusetts 02115, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Harvard Institutes of Human Genetics, Harvard Medical School, Beth Israel Deaconess Medical Center, 4 Blackfan Circle, Boston, Massachusetts 02115</wicri:regionArea><wicri:noRegion>Massachusetts 02115</wicri:noRegion></affiliation></author><author><name sortKey="Hong, Y K" sort="Hong, Y K" uniqKey="Hong Y" first="Y K" last="Hong">Y K Hong</name></author><author><name sortKey="Ontiveros, S D" sort="Ontiveros, S D" uniqKey="Ontiveros S" first="S D" last="Ontiveros">S D Ontiveros</name></author><author><name sortKey="Egholm, M" sort="Egholm, M" uniqKey="Egholm M" first="M" last="Egholm">M. 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Chen</name><affiliation wicri:level="1"><nlm:affiliation>Harvard Institutes of Human Genetics, Harvard Medical School, Beth Israel Deaconess Medical Center, 4 Blackfan Circle, Boston, Massachusetts 02115, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Harvard Institutes of Human Genetics, Harvard Medical School, Beth Israel Deaconess Medical Center, 4 Blackfan Circle, Boston, Massachusetts 02115</wicri:regionArea><wicri:noRegion>Massachusetts 02115</wicri:noRegion></affiliation></author><author><name sortKey="Hong, Y K" sort="Hong, Y K" uniqKey="Hong Y" first="Y K" last="Hong">Y K Hong</name></author><author><name sortKey="Ontiveros, S D" sort="Ontiveros, S D" uniqKey="Ontiveros S" first="S D" last="Ontiveros">S D Ontiveros</name></author><author><name sortKey="Egholm, M" sort="Egholm, M" uniqKey="Egholm M" first="M" last="Egholm">M. Egholm</name></author><author><name sortKey="Strauss, W M" sort="Strauss, W M" uniqKey="Strauss W" first="W M" last="Strauss">W M Strauss</name></author></analytic><series><title level="j">Mammalian genome : official journal of the International Mammalian Genome Society</title><idno type="ISSN">0938-8990</idno><imprint><date when="1999" type="published">1999</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Autoantigens</term><term>Base Pairing</term><term>Centromere Protein B</term><term>Chromosomal Proteins, Non-Histone (genetics)</term><term>DNA Primers</term><term>DNA-Binding Proteins</term><term>Humans</term><term>In Situ Hybridization, Fluorescence (methods)</term><term>Mice</term><term>Mice, Inbred C57BL</term><term>Peptide Nucleic Acids</term><term>Repetitive Sequences, Nucleic Acid</term><term>Sensitivity and Specificity</term><term>Species Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Acides nucléiques peptidiques</term><term>Amorces ADN</term><term>Animaux</term><term>Appariement de bases</term><term>Autoantigènes</term><term>Humains</term><term>Protéine B du centromère</term><term>Protéines chromosomiques nonhistones (génétique)</term><term>Protéines de liaison à l'ADN</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Souris de lignée C57BL</term><term>Spécificité d'espèce</term><term>Séquences répétées d'acides nucléiques</term><term>Technique FISH ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Chromosomal Proteins, Non-Histone</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Autoantigens</term><term>Centromere Protein B</term><term>DNA Primers</term><term>DNA-Binding Proteins</term><term>Peptide Nucleic Acids</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines chromosomiques nonhistones</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>In Situ Hybridization, Fluorescence</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Pairing</term><term>Humans</term><term>Mice</term><term>Mice, Inbred C57BL</term><term>Repetitive Sequences, Nucleic Acid</term><term>Sensitivity and Specificity</term><term>Species Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Acides nucléiques peptidiques</term><term>Amorces ADN</term><term>Animaux</term><term>Appariement de bases</term><term>Autoantigènes</term><term>Humains</term><term>Protéine B du centromère</term><term>Protéines de liaison à l'ADN</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Souris de lignée C57BL</term><term>Spécificité d'espèce</term><term>Séquences répétées d'acides nucléiques</term><term>Technique FISH</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The satellite repeat structure of the mammalian centromere contains the CENP-B protein binding site. Using the peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH), we show by direct PNA-DNA binding that all detectable CENP-B sites in a mammalian genome might have the same sequence. Two species-specific PNA 17-mers, pMm and pMc, were identified from CENP-B binding sites of Mus musculus and M. caroli, respectively. Fluorescence in situ hybridization confirmed that pMc hybridized to M. caroli centromeres only; however, pMm cross-hybridized to M. musculus and human centromeres. By using a series of CENP-B PNA 17-mers containing 1, 2, 3, 5, and 7 base-pair mismatches to their DNA counterparts, we further demonstrate that PNA-FISH can discriminate between two CENP-B DNA sequences that differ by a single base-pair in mouse and human centromeres, suggesting the degree of conservation of CENP-B sequences throughout the genome. In comparison with DNA oligonucleotides, PNA oligomers demonstrate the higher sequence specificity, improved stability, reproducibility, and lower background. Therefore, PNA oligomers have significant advantages over DNA oligonucleotide probes in analyzing microsatellites in a genome.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Egholm, M" sort="Egholm, M" uniqKey="Egholm M" first="M" last="Egholm">M. Egholm</name><name sortKey="Hong, Y K" sort="Hong, Y K" uniqKey="Hong Y" first="Y K" last="Hong">Y K Hong</name><name sortKey="Ontiveros, S D" sort="Ontiveros, S D" uniqKey="Ontiveros S" first="S D" last="Ontiveros">S D Ontiveros</name><name sortKey="Strauss, W M" sort="Strauss, W M" uniqKey="Strauss W" first="W M" last="Strauss">W M Strauss</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Chen, C" sort="Chen, C" uniqKey="Chen C" first="C" last="Chen">C. 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