The Role of Transcription in the Activation of a Drosophila Amplification Origin
The mechanisms that underlie metazoan DNA replication initiation, especially the connection between transcription and replication origin activation, are not well understood. To probe the role of transcription in origin activation, we exploited a specific replication origin in Drosophila melanogaster...
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| Format: | Article |
| Language: | en_US |
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Genetics Society of America, The
2015
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| Online Access: | http://hdl.handle.net/1721.1/96690 https://orcid.org/0000-0002-7934-111X https://orcid.org/0000-0002-7580-3399 |
| _version_ | 1826215654578978816 |
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| author | Hua, Brian Li, Sharon Orr-Weaver, Terry |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Hua, Brian Li, Sharon Orr-Weaver, Terry |
| author_sort | Hua, Brian |
| collection | MIT |
| description | The mechanisms that underlie metazoan DNA replication initiation, especially the connection between transcription and replication origin activation, are not well understood. To probe the role of transcription in origin activation, we exploited a specific replication origin in Drosophila melanogaster follicle cells, ori62, which coincides with the yellow-g2 transcription unit and exhibits transcription-dependent origin firing. Within a 10-kb genomic fragment that contains ori62 and is sufficient for amplification, RNA-sequencing analysis revealed that all detected RNAs mapped solely to the yellow-g2 gene. To determine whether transcription is required in cis for ori62 firing, we generated a set of tagged yellow-g2 transgenes in which we could prevent local transcription across ori62 by deletions in the yellow-g2 promoter. Surprisingly, inhibition of yellow-g2 transcription by promoter deletions did not affect ori62 firing. Our results reveal that transcription in cis is not required for ori62 firing, raising the possibility that a trans-acting factor is required specifically for the activation of ori62. This finding illustrates that a diversity of mechanisms can be used in the regulation of metazoan DNA replication initiation. |
| first_indexed | 2024-09-23T16:37:21Z |
| format | Article |
| id | mit-1721.1/96690 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:37:21Z |
| publishDate | 2015 |
| publisher | Genetics Society of America, The |
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| spelling | mit-1721.1/966902022-09-29T20:21:58Z The Role of Transcription in the Activation of a Drosophila Amplification Origin Hua, Brian Li, Sharon Orr-Weaver, Terry Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Orr-Weaver, Terry L. Hua, Brian The mechanisms that underlie metazoan DNA replication initiation, especially the connection between transcription and replication origin activation, are not well understood. To probe the role of transcription in origin activation, we exploited a specific replication origin in Drosophila melanogaster follicle cells, ori62, which coincides with the yellow-g2 transcription unit and exhibits transcription-dependent origin firing. Within a 10-kb genomic fragment that contains ori62 and is sufficient for amplification, RNA-sequencing analysis revealed that all detected RNAs mapped solely to the yellow-g2 gene. To determine whether transcription is required in cis for ori62 firing, we generated a set of tagged yellow-g2 transgenes in which we could prevent local transcription across ori62 by deletions in the yellow-g2 promoter. Surprisingly, inhibition of yellow-g2 transcription by promoter deletions did not affect ori62 firing. Our results reveal that transcription in cis is not required for ori62 firing, raising the possibility that a trans-acting factor is required specifically for the activation of ori62. This finding illustrates that a diversity of mechanisms can be used in the regulation of metazoan DNA replication initiation. National Institutes of Health (U.S.) (Grant GM57960) 2015-04-21T18:55:25Z 2015-04-21T18:55:25Z 2014-10 2014-08 Article http://purl.org/eprint/type/JournalArticle 2160-1836 http://hdl.handle.net/1721.1/96690 Hua, B. L., S. Li, and T. L. Orr-Weaver. “The Role of Transcription in the Activation of a Drosophila Amplification Origin.” G3: Genes-Genomes-Genetics 4, no. 12 (October 14, 2014): 2403–2408. https://orcid.org/0000-0002-7934-111X https://orcid.org/0000-0002-7580-3399 en_US http://dx.doi.org/10.1534/g3.114.014050 G3: Genes-Genomes-Genetics Creative Commons Attribution Unported License http://creativecommons.org/licenses/by/3.0/ application/pdf Genetics Society of America, The G3 |
| spellingShingle | Hua, Brian Li, Sharon Orr-Weaver, Terry The Role of Transcription in the Activation of a Drosophila Amplification Origin |
| title | The Role of Transcription in the Activation of a Drosophila Amplification Origin |
| title_full | The Role of Transcription in the Activation of a Drosophila Amplification Origin |
| title_fullStr | The Role of Transcription in the Activation of a Drosophila Amplification Origin |
| title_full_unstemmed | The Role of Transcription in the Activation of a Drosophila Amplification Origin |
| title_short | The Role of Transcription in the Activation of a Drosophila Amplification Origin |
| title_sort | role of transcription in the activation of a drosophila amplification origin |
| url | http://hdl.handle.net/1721.1/96690 https://orcid.org/0000-0002-7934-111X https://orcid.org/0000-0002-7580-3399 |
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