Identification of transcription factors coupling the cell cycle machinery with environmental and developmental signals

In the recent years many studies contributed to our understanding of the mechanisms that drives the cell cycle machinery. Studies performed in yeast, animals, worms, flies and plants revealed that despite the evolutionary distance between these species, an universal picture can be drawn on how the basic cell cycle machinery is regulated. However, in spite of their highly conserved cell cycle machinery, it is remarkable how plants and animals have integrated the control of cell cycle differently into their specific developmental programs. In contrast to animals, plants develop mostly post-embryonic, which is characterized by continuous growth and organ formation during their entire life-span. This developmental style relies on the existence of stem cell niches within the root and shoot apical meristems, continuously supplying new cells. Next to this, plants are able to form organs de novo, like lateral roots, requiring cell cycle reactivation within already differentiated cells. Additionally, besides cell proliferation, endoreduplication plays as well an important role during different developmental processes. The plant body shows an amazing flexibility rendering them the ability to cope with different environmental and developmental signals. However, although different reports describe the influence of different environmental and developmental cues on cell cycle progression and endoreduplication, we have currently only limited knowledge on how these signals connect to the core cell cycle machinery. The aim of this project was to gain insight in how these intrinsic and extrinsic signals are integrated with the regulation of the cell cycle machinery. Although different core cell cycle genes display developmental and cell cycle-phase dependent transcriptional regulation, it is intriguing how little is known on their transcriptional regulators. Therefore the work was focused on determining new transcriptional regulators of core cell cycle genes and to try to connect these with specific environmental or developmental processes

Arabidopsis ULTRAVIOLET-B-INSENSITIVE4 maintains cell division activity by temporal inhibition of the anaphase-promoting complex/cyclosome

The anaphase-promoting complex/cyclosome (APC/C) is a multisubunit ubiquitin ligase that regulates progression through the cell cycle by marking key cell division proteins for destruction. To ensure correct cell cycle progression, accurate timing of APC/C activity is important, which is obtained through its association with both activating and inhibitory subunits. However, although the APC/C is highly conserved among eukaryotes, no APC/C inhibitors are known in plants. Recently, we have identified ULTRAVIOLET-B-INSENSITIVE4 (UVI4) as a plant-specific component of the APC/C. Here, we demonstrate that UVI4 uses conserved APC/C interaction motifs to counteract the activity of the CELL CYCLE SWITCH52 A1 (CCS52A1) activator subunit, inhibiting the turnover of the A-type cyclin CYCA2;3. UVI4 is expressed in an S phase-dependent fashion, likely through the action of E2F transcription factors. Correspondingly, uvi4 mutant plants failed to accumulate CYCA2; 3 during the S phase and prematurely exited the cell cycle, triggering the onset of the endocycle. We conclude that UVI4 regulates the temporal inactivation of APC/C during DNA replication, allowing CYCA2;3 to accumulate above the level required for entering mitosis, and thereby regulates the meristem size and plant growth rate

Auxin-dependent cell cycle reactivation through transcriptional regulation of Arabidopsis E2Fa by lateral organ boundary proteins

Multicellular organisms depend on cell production, cell fate specification, and correct patterning to shape their adult body. In plants, auxin plays a prominent role in the timely coordination of these different cellular processes. A well-studied example is lateral root initiation, in which auxin triggers founder cell specification and cell cycle activation of xylem pole-positioned pericycle cells. Here, we report that the E2Fa transcription factor of Arabidopsis thaliana is an essential component that regulates the asymmetric cell division marking lateral root initiation. Moreover, we demonstrate that E2Fa expression is regulated by the LATERAL ORGAN BOUNDARY DOMAIN18/LATERAL ORGAN BOUNDARY DOMAIN33 (LBD18/LBD33) dimer that is, in turn, regulated by the auxin signaling pathway. LBD18/LBD33 mediates lateral root organogenesis through E2Fa transcriptional activation, whereas E2Fa expression under control of the LBD18 promoter eliminates the need for LBD18. Besides lateral root initiation, vascular patterning is disrupted in E2Fa knockout plants, similarly as it is affected in auxin signaling and lbd mutants, indicating that the transcriptional induction of E2Fa through LBDs represents a general mechanism for auxin-dependent cell cycle activation. Our data illustrate how a conserved mechanism driving cell cycle entry has been adapted evolutionarily to connect auxin signaling with control of processes determining plant architecture

Comparative Transcriptome Atlases Reveal Altered Gene Expression Modules between Two Cleomaceae C-3 and C-4 Plant Species

Külahoglu C, Denton AK, Sommer M, et al. Comparative Transcriptome Atlases Reveal Altered Gene Expression Modules between Two Cleomaceae C-3 and C-4 Plant Species. Plant Cell. 2014;26(8):3243-3260.C-4 photosynthesis outperforms the ancestral C-3 state in a wide range of natural and agro-ecosystems by affording higher water-use and nitrogen-use efficiencies. It therefore represents a prime target for engineering novel, high-yielding crops by introducing the trait into C-3 backgrounds. However, the genetic architecture of C-4 photosynthesis remains largely unknown. To define the divergence in gene expression modules between C-3 and C-4 photosynthesis during leaf ontogeny, we generated comprehensive transcriptome atlases of two Cleomaceae species, Gynandropsis gynandra (C-4) and Tarenaya hassleriana (C-3), by RNA sequencing. Overall, the gene expression profiles appear remarkably similar between the C-3 and C-4 species. We found that known C-4 genes were recruited to photosynthesis from different expression domains in C-3, including typical housekeeping gene expression patterns in various tissues as well as individual heterotrophic tissues. Furthermore, we identified a structure-related module recruited from the C-3 root. Comparison of gene expression patterns with anatomy during leaf ontogeny provided insight into genetic features of Kranz anatomy. Altered expression of developmental factors and cell cycle genes is associated with a higher degree of endoreduplication in enlarged C-4 bundle sheath cells. A delay in mesophyll differentiation apparent both in the leaf anatomy and the transcriptome allows for extended vein formation in the C-4 leaf

Protein complex stoichiometry and expression dynamics of transcription factors modulate stem cell division

11 Pág.Stem cells divide and differentiate to form all of the specialized cell types in a multicellular organism. In the Arabidopsis root, stem cells are maintained in an undifferentiated state by a less mitotically active population of cells called the quiescent center (QC). Determining how the QC regulates the surrounding stem cell initials, or what makes the QC fundamentally different from the actively dividing initials, is important for understanding how stem cell divisions are maintained. Here we gained insight into the differences between the QC and the cortex endodermis initials (CEI) by studying the mobile transcription factor SHORTROOT (SHR) and its binding partner SCARECROW (SCR). We constructed an ordinary differential equation model of SHR and SCR in the QC and CEI which incorporated the stoichiometry of the SHR-SCR complex as well as upstream transcriptional regulation of SHR and SCR. Our model prediction, coupled with experimental validation, showed that high levels of the SHR-SCR complex are associated with more CEI division but less QC division. Furthermore, our model prediction allowed us to propose the putative upstream SHR regulators SEUSS and WUSCHEL-RELATED HOMEOBOX 5 and to experimentally validate their roles in QC and CEI division. In addition, our model established the timing of QC and CEI division and suggests that SHR repression of QC division depends on formation of the SHR homodimer. Thus, our results support that SHR-SCR protein complex stoichiometry and regulation of SHR transcription modulate the division timing of two different specialized cell types in the root stem cell niche.This work was supported by the NSF Graduate Research Fellowship Program (DGE-1252376, to N.M.C. and A.P.F.). Research in the R. Simon lab was funded by the Deutsche Forschungsge-meinschaft (Si947/10 and an Alexander von Humboldt Foundation fellowship, to B.B.). This work was also supported by a grant from the Ministerio de Economía y Competitividad of Spain and European Regional Development Fund (BFU2016-80315-P, to M.A.M.-R.). E.B.A. is supported by Ayudante de Investigacion contract PEJ-2017-AI/BIO-7360 from Comunidad Madrid. S.G.Z. was supported by the HHMI and by a grant from the NIH (GM118036). Research in the K.L.G. lab was funded by NSF Grant 1243945. The R. Sozzani lab is supported by an NSF CAREER grant (MCB-1453130) and the North Carolina Agricultural & Life Sciences Research Foundation at North Carolina State University’s College of Agricultural and Life Sciences.Peer reviewe

Transcriptional control of the cell cycle

Cell division is a highly coordinated process. In the last decades, many plant cell cycle regulators have been identified. Strikingly, only a few transcriptional regulators are known, although a significant amount of the genome is transcribed in a cell cycle phase-dependent manner. E2F-DP transcription factors and three repeat MYB proteins are responsible for the expression of genes at the G1-to-S and G2-to-M transition, respectively. However, these two mechanisms cannot explain completely the transcriptional regulation seen during the cell cycle. Correspondingly, several new transcriptional regulators have been characterized, stressing the importance of transcriptional control during the cell cycle

Screening for novel biocontrol agents applicable in plant disease management - A review

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The E2F transcription factor family regulates CENH3 expression in Arabidopsis thaliana

To elucidate the epigenetic maintenance mechanism for functional plant centromeres, we studied transcriptional regulation of the centromere-specific histone H3 variant CENH3 in Arabidopsis thaliana. We focused on the structure and activity of the CENH3 promoter (CENH3pro) and its regulation by E2F transcription factors. Use of CENH3pro::GUS reporter gene constructs showed that CENH3pro is active in dividing tissues, and that full expression in root meristems depends on intragenic regulatory elements within the second intron. Chromatin immunoprecipitation identified CENH3 as an E2F target gene. Transient co-expression of a CENH3pro:: GUS reporter gene construct with various E2F transcription factors in A. thaliana protoplasts showed that E2Fa and E2Fb (preferentially with dimerization protein DPb) activate CENH3pro. Stable overexpression of E2Fa and E2Fb increased the CENH3 transcript level in planta, whereas over-expression of E2Fc decreased the CENH3 transcript level. Surprisingly, mutation of the two E2F binding sites of CENH3pro, in particular the more upstream one (E2F2), caused an increase in CENH3pro activity, indicating E2F-dependent transcriptional repression. CENH3pro repression may be triggered by the interplay of typical and atypical E2Fs in a cell cycle-dependent manner, and/or by interaction of typical E2Fs with retinoblastoma-related (RBR) protein. We speculate that E2Fs are involved in differential transcriptional regulation of CENH3 versus H3, as H3 promoters lack E2F binding motifs. E2F binding motifs are also present in human and Drosophila CENH3pro regions, thus cell cycle-dependent transcriptional regulation of CENH3 may be highly conserved