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The intersection of DNA replication with antisense 3' RNA processing in Arabidopsis FLC chromatin silencing.


ABSTRACT: How noncoding transcription influences chromatin states is still unclear. The Arabidopsis floral repressor gene FLC is quantitatively regulated through an antisense-mediated chromatin silencing mechanism. The FLC antisense transcripts form a cotranscriptional R-loop that is dynamically resolved by RNA 3' processing factors (FCA and FY), and this is linked to chromatin silencing. Here, we investigate this silencing mechanism and show, using single-molecule DNA fiber analysis, that FCA and FY are required for unimpeded replication fork progression across the Arabidopsis genome. We then employ the chicken DT40 cell line system, developed to investigate sequence-dependent replication and chromatin inheritance, and find that FLC R-loop sequences have an orientation-dependent ability to stall replication forks. These data suggest a coordination between RNA 3' processing of antisense RNA and replication fork progression in the inheritance of chromatin silencing at FLC.

SUBMITTER: Baxter CL 

PROVIDER: S-EPMC8285979 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature

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The intersection of DNA replication with antisense 3' RNA processing in <i>Arabidopsis FLC</i> chromatin silencing.

Baxter Colette L CL   Šviković Saša S   Sale Julian E JE   Dean Caroline C   Costa Silvia S  

Proceedings of the National Academy of Sciences of the United States of America 20210701 28


How noncoding transcription influences chromatin states is still unclear. The <i>Arabidopsis</i> floral repressor gene <i>FLC</i> is quantitatively regulated through an antisense-mediated chromatin silencing mechanism. The <i>FLC</i> antisense transcripts form a cotranscriptional R-loop that is dynamically resolved by RNA 3' processing factors (FCA and FY), and this is linked to chromatin silencing. Here, we investigate this silencing mechanism and show, using single-molecule DNA fiber analysis,  ...[more]

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