The shoot apical meristem (SAM), which produces all the aerial parts of the plant, plays a major role in the establish- ment of plant architecture. In many higher plants, the SAM has a stereotypic organization in zo...The shoot apical meristem (SAM), which produces all the aerial parts of the plant, plays a major role in the establish- ment of plant architecture. In many higher plants, the SAM has a stereotypic organization in zones with different func- tions. At the summit of the dome-shaped meristem is a popu- lation of stem cells (called the central zone), required for meristem maintenance. Part of the cells produced by the central zone will enter a differentiation pathway and their descendants will serve as founders for the stems and lateral organs at the meristem periphery (or peripheral zone). During the last decades, much of the research on the SAM has focused on three major signaling cascades required for its function:展开更多
Auxin is a key hormone performing a wealth of functions throughout the life cycle of plants. It acts largely by regulating genes at the transcriptional level through a family of transcription factors called auxin resp...Auxin is a key hormone performing a wealth of functions throughout the life cycle of plants. It acts largely by regulating genes at the transcriptional level through a family of transcription factors called auxin response factors (ARFs). Even though all ARF monomers analyzed so far bind a similar DNA sequence, there is evidence that ARFs differ in their target genomic regions and regulated genes. Here, we report the use of position weight matrices (PWMs) to model ARF DNA binding specificity based on published DNA affinity purification sequencing (DAP-seq) data. We found that the genome binding of two ARFs (ARF2 and ARF5/ Monopteros [MP]) differ largely because these two factors have different preferred ARF binding site (ARFbs) arrangements (orientation and spacing). We illustrated why PWMs are more versatile to reliably identify ARFbs than the widely used consensus sequences and demonstrated their power with biochemical experiments in the identification of the regulatory regions o1IAA19, an well-characterized auxin-responsive gene. Finally, we combined gene regulation by auxin with ARF-bound regions and identified specific ARFbs configurations that are over-represented in auxin-upregulated genes, thus deciphering the ARFbs syntax functional for regulation. Our study provides a general method to exploit the potential of genome-wide DNA binding assays and to decode gene regulation.展开更多
Pioneer transcription factors(TFs)are a special category of TFs with the capacity to bind to closed chromatin regions in which DNA is wrapped around histones and may be highly methylated.Subsequently,pioneer TFs are a...Pioneer transcription factors(TFs)are a special category of TFs with the capacity to bind to closed chromatin regions in which DNA is wrapped around histones and may be highly methylated.Subsequently,pioneer TFs are able to modify the chromatin state to initiate gene expression.In plants,LEAFY(LFY)is a master floral regulator and has been suggested to act as a pioneer TF in Arabidopsis.Here,we demonstrate that LFY is able to bind both methylated and non-methylated DNA using a combination of in vitro genomewide binding experiments and structural modeling.Comparisons between regions bound by LFY in vivo and chromatin accessibility data suggest that a subset of LFY bound regions is occupied by nucleosomes.We confirm that LFY is able to bind nucleosomal DNA in vitro using reconstituted nucleosomes.Finally,we show that constitutive LFY expression in seedling tissues is sufficient to induce chromatin accessibility in the LFY direct target genes APETALA1 and AGAMOUS.Taken together,our study suggests that LFY possesses key pioneer TF features that contribute to launching the floral gene expression program.展开更多
文摘The shoot apical meristem (SAM), which produces all the aerial parts of the plant, plays a major role in the establish- ment of plant architecture. In many higher plants, the SAM has a stereotypic organization in zones with different func- tions. At the summit of the dome-shaped meristem is a popu- lation of stem cells (called the central zone), required for meristem maintenance. Part of the cells produced by the central zone will enter a differentiation pathway and their descendants will serve as founders for the stems and lateral organs at the meristem periphery (or peripheral zone). During the last decades, much of the research on the SAM has focused on three major signaling cascades required for its function:
文摘Auxin is a key hormone performing a wealth of functions throughout the life cycle of plants. It acts largely by regulating genes at the transcriptional level through a family of transcription factors called auxin response factors (ARFs). Even though all ARF monomers analyzed so far bind a similar DNA sequence, there is evidence that ARFs differ in their target genomic regions and regulated genes. Here, we report the use of position weight matrices (PWMs) to model ARF DNA binding specificity based on published DNA affinity purification sequencing (DAP-seq) data. We found that the genome binding of two ARFs (ARF2 and ARF5/ Monopteros [MP]) differ largely because these two factors have different preferred ARF binding site (ARFbs) arrangements (orientation and spacing). We illustrated why PWMs are more versatile to reliably identify ARFbs than the widely used consensus sequences and demonstrated their power with biochemical experiments in the identification of the regulatory regions o1IAA19, an well-characterized auxin-responsive gene. Finally, we combined gene regulation by auxin with ARF-bound regions and identified specific ARFbs configurations that are over-represented in auxin-upregulated genes, thus deciphering the ARFbs syntax functional for regulation. Our study provides a general method to exploit the potential of genome-wide DNA binding assays and to decode gene regulation.
基金supported by the ANR-DFG project Flopinet(ANR-16-CE92-0023-01)to C.Z.and F.P.,and GRALa program from the Chemistry Biology Health(CBH)Graduate School of University Grenoble Alpes(ANR-17-EURE-0003)to C.Z.,F.P.,and A.S.
文摘Pioneer transcription factors(TFs)are a special category of TFs with the capacity to bind to closed chromatin regions in which DNA is wrapped around histones and may be highly methylated.Subsequently,pioneer TFs are able to modify the chromatin state to initiate gene expression.In plants,LEAFY(LFY)is a master floral regulator and has been suggested to act as a pioneer TF in Arabidopsis.Here,we demonstrate that LFY is able to bind both methylated and non-methylated DNA using a combination of in vitro genomewide binding experiments and structural modeling.Comparisons between regions bound by LFY in vivo and chromatin accessibility data suggest that a subset of LFY bound regions is occupied by nucleosomes.We confirm that LFY is able to bind nucleosomal DNA in vitro using reconstituted nucleosomes.Finally,we show that constitutive LFY expression in seedling tissues is sufficient to induce chromatin accessibility in the LFY direct target genes APETALA1 and AGAMOUS.Taken together,our study suggests that LFY possesses key pioneer TF features that contribute to launching the floral gene expression program.