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Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain
The maize B centromere has been used as a model for centromere epigenetics and as the basis for building artificial chromosomes. However, there are no sequence resources for this important centromere. Here we used transposon display for the centromere-specific retroelement CRM2 to identify a collect...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Genetics Society of America
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4683656/ https://www.ncbi.nlm.nih.gov/pubmed/26511496 http://dx.doi.org/10.1534/g3.115.022889 |
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author | Ellis, Nathanael A. Douglas, Ryan N. Jackson, Caroline E. Birchler, James A. Dawe, R. Kelly |
author_facet | Ellis, Nathanael A. Douglas, Ryan N. Jackson, Caroline E. Birchler, James A. Dawe, R. Kelly |
author_sort | Ellis, Nathanael A. |
collection | PubMed |
description | The maize B centromere has been used as a model for centromere epigenetics and as the basis for building artificial chromosomes. However, there are no sequence resources for this important centromere. Here we used transposon display for the centromere-specific retroelement CRM2 to identify a collection of 40 sequence tags that flank CRM2 insertion points on the B chromosome. These were confirmed to lie within the centromere by assaying deletion breakpoints from centromere misdivision derivatives (intracentromere breakages caused by centromere fission). Markers were grouped together on the basis of their association with other markers in the misdivision series and assembled into a pseudocontig containing 10.1 kb of sequence. To identify sequences that interact directly with centromere proteins, we carried out chromatin immunoprecipitation using antibodies to centromeric histone H3 (CENH3), a defining feature of functional centromeric sequences. The CENH3 chromatin immunoprecipitation map was interpreted relative to the known transmission rates of centromere misdivision derivatives to identify a centromere core domain spanning 33 markers. A subset of seven markers was mapped in additional B centromere misdivision derivatives with the use of unique primer pairs. A derivative previously shown to have no canonical centromere sequences (Telo3-3) lacks these core markers. Our results provide a molecular map of the B chromosome centromere and identify key sequences within the map that interact directly with centromeric histone H3. |
format | Online Article Text |
id | pubmed-4683656 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Genetics Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-46836562015-12-18 Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain Ellis, Nathanael A. Douglas, Ryan N. Jackson, Caroline E. Birchler, James A. Dawe, R. Kelly G3 (Bethesda) Investigations The maize B centromere has been used as a model for centromere epigenetics and as the basis for building artificial chromosomes. However, there are no sequence resources for this important centromere. Here we used transposon display for the centromere-specific retroelement CRM2 to identify a collection of 40 sequence tags that flank CRM2 insertion points on the B chromosome. These were confirmed to lie within the centromere by assaying deletion breakpoints from centromere misdivision derivatives (intracentromere breakages caused by centromere fission). Markers were grouped together on the basis of their association with other markers in the misdivision series and assembled into a pseudocontig containing 10.1 kb of sequence. To identify sequences that interact directly with centromere proteins, we carried out chromatin immunoprecipitation using antibodies to centromeric histone H3 (CENH3), a defining feature of functional centromeric sequences. The CENH3 chromatin immunoprecipitation map was interpreted relative to the known transmission rates of centromere misdivision derivatives to identify a centromere core domain spanning 33 markers. A subset of seven markers was mapped in additional B centromere misdivision derivatives with the use of unique primer pairs. A derivative previously shown to have no canonical centromere sequences (Telo3-3) lacks these core markers. Our results provide a molecular map of the B chromosome centromere and identify key sequences within the map that interact directly with centromeric histone H3. Genetics Society of America 2015-10-26 /pmc/articles/PMC4683656/ /pubmed/26511496 http://dx.doi.org/10.1534/g3.115.022889 Text en Copyright © 2015 Ellis et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Investigations Ellis, Nathanael A. Douglas, Ryan N. Jackson, Caroline E. Birchler, James A. Dawe, R. Kelly Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain |
title | Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain |
title_full | Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain |
title_fullStr | Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain |
title_full_unstemmed | Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain |
title_short | Generation of a Maize B Centromere Minimal Map Containing the Central Core Domain |
title_sort | generation of a maize b centromere minimal map containing the central core domain |
topic | Investigations |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4683656/ https://www.ncbi.nlm.nih.gov/pubmed/26511496 http://dx.doi.org/10.1534/g3.115.022889 |
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