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A Structural Bisulfite Assay to Identify DN Cruciforms

Molecular Plant 2016年第9期9卷 1328-1336页

作者： Matthew Gentry lars hennig Department of Plant Biology Uppsala BioCenter Swedish University of Agricultural Sciences and Linnean Center for Plant Biology 75007 Uppsala Sweden

In the half century since the discovery of the double-helix structure of DNA, it has become increasingly clear that DNA functionality is based on much more than its sequence in a double-helical structure. Further advances have highlighted the importance of additional aspects of DNA structure： its packaging in the higher order chromatin structure, positioning of nucleosomes along the DNA, and the occurrence of non-helical DNA structures. Of these, the latter has been problematic to prove empirically. Here, we describe a method that uses non-denaturing bisulfite sequencing on isolated Arabidopsis thaliana nuclei to determine the location of cytosines positioned outside the double helix as a result of non-B-form DNA structures. We couple this with computational methods and S1 nuclease digest to reliably identify stable, non-B-form, cruciform structures. This enables us to identify a palindrome in the promoter of FLOWERING LOCUS T that forms a stable non-B-form structure. The stronger conservation of the ability to form a nonhelical secondary structure than of the sequence suggests that this structure is biologically relevant.

关键词： DNA structure cruciform DNA genome organization

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Lost Memories of Winter： Breaking the FLC Silence

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Molecular Plant 2017年第12期10卷 1477-1479页

作者： Miyuki Nakamura Iva Mozgova lars hennig Department of Plant Biology Uppsala BioCenter Swedish University of Agricultural Sciences and Linnean Center for Plant Biology 75007 Uppsala Sweden Institute of Microbiology of the Czech Academy of Sciences Centre Algatech Opatovický mlýn 37981 Třeboň Czech Republic Faculty of Science University of South Bohemia in Ceske Budejovice 37005 Ceske Budejovice Czech Republic

Vernalization, the acquisition of competence to flower after extended cold treatment, has puzzled researchers throughout the last century. Great progress in understanding the molecular mechanisms of vernalization in Arabidopsis has been made in the course of the last 20 years, placing the flowering repressor- encoding gene FLOWERING LOCUS C （FLC） in the limelight （Bouch6 et al., 2017）. In vernalization-dependent accessions, FRIGIDA （FRI） ensures strong expression of FLC that delays flowering, and extended exposure to low temperatures such as during winter leads to stable repression of FLC, which in turn clears the way for induction of flowering.

关键词： FLOWERING-LOCUS-C TRANSCRIPTION FACTOR VERNALIZATION INHERITANCE ARABIDOPSIS EXPRESSION

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Arabidopsis MSI1 Is Required for Negative Regulation of the Response to Drought Stress

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Molecular Plant 2009年第4期2卷 675-687页

作者： Cristina Alexandre Yvonne Moller-Steinbach Nicole Sch6nrock Wilhelm Gruissem lars hennig Department of Biology and Zurich-Basel Plant Science Center ETH Zurich CH-8092 Zurich Switzerland Present address: Brain and Mind Research Institute University of Sydney 100 Mallett St Camperdown NSW 2050 Australia Department of Biology ETH Zurich LFW E17 CH-8092 Zurich Switzerland

Arabidopsis MSI1 has fundamental functions in plant development. MSI1 is a subunit of Polycomb group protein complexes and Chromatin assembly factor 1, and it interacts with the Retinoblastoma-related protein 1. Altered levels of MSI1 result in pleiotropic phenotypes, reflecting the complexity of MSI1 protein functions. In order to uncover additional functions of MSI1, we performed transcriptional profiling of wild-type and plants with highly reduced MSI1 levels （msil-cs）. Surprisingly, the known functions of MSI1 could only account for a minor part of the transcriptional changes in msil-cs plants. One of the most striking unexpected observations was the up-regulation of a subset of ABA-responsive genes eliciting the response to drought and salt stress. We report that MSI1 can bind to the chromatin of the drought-inducible downstream target RD20 and suggest a new role for MSI1 in the negative regulation of the Arabidopsis drought-stress response.

关键词： Abiotic/environmental stress water relations chromatin structure and remodeling transcriptome analysis development.

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