Mediator subunit MED31 is required for radial patterning of Arabidopsis roots

Stem cell specification in multicellular organisms relies on the precise spatiotemporal control of RNA polymerase II (Pol II)-dependent gene transcription, in which the evolutionarily conserved Mediator coactivator complex plays an essential role. In Arabidopsis thaliana, SHORTROOT (SHR) and SCARECROW (SCR) orchestrate a transcriptional program that determines the fate and asymmetrical divisions of stem cells generating the root ground tissue. The mechanism by which SHR/SCR relays context-specific regulatory signals to the Pol II general transcription machinery is unknown. Here, we report the role of Mediator in controlling the spatiotemporal transcriptional output of SHR/SCR during asymmetrical division of stem cells and ground tissue patterning. The Mediator subunit MED31 interacted with SCR but not SHR. Reduction of MED31 disrupted the spatiotemporal activation of CYCLIND6;1 (CYCD6;1), leading to defective asymmetrical division of stem cells generating ground tissue. MED31 was recruited to the promoter of CYCD6;1 in an SCR-dependent manner. MED31 was involved in the formation of a dynamic MED31/SCR/SHR ternary complex through the interface protein SCR. We demonstrate that the relative protein abundance of MED31 and SHR in different cell types regulates the dynamic formation of the ternary complex, which provides a tunable switch to strictly control the spatiotemporal transcriptional output. This study provides valuable clues to understand the mechanism by which master transcriptional regulators control organ patterning.

Pax6 is required to regulate the cell cycle and the rate of progression from symmetrical to asymmetrical division in mammalian cortical progenitors

In the proliferative zone of the developing cerebral cortex, multipotential progenitors predominate early in development and divide to increase the progenitor pool. As corticogenesis progresses, proportionately fewer progenitors are produced and, instead, cell divisions yield higher numbers of postmitotic neurones or glial cells. As the switch from the generation of progenitors to that of differentiated cells occurs, the orientation of cell division alters from predominantly symmetrical to predominantly asymmetrical. It has been hypothesised that symmetrical divisions expand the progenitor pool, whereas asymmetrical divisions generate postmitotic cells, although this remains to be proved. The molecular mechanisms regulating these processes are poorly understood. The transcription factor Pax6 is highly expressed in the cortical proliferative zone and there are morphological defects in the Pax6Sey/Sey (Pax6 null) cortex, but little is known about the principal cellular functions of Pax6 in this region. We have analysed the cell-cycle kinetics, the progenitor cleavage orientation and the onset of expression of differentiation markers in Pax6Sey/Sey cortical cells in vivo and in vitro. We showed that, early in corticogenesis at embryonic day (E) 12.5, the absence of Pax6 accelerated cortical development in vivo, shortening the cell cycle and the time taken for the onset of expression of neural-specific markers. This also occurred in dissociated culture of isolated cortical cells, indicating that the changes were intrinsic to the cortical cells. From E12.5 to E15.5, proportions of asymmetrical divisions increased more rapidly in mutant than in wild-type embryos. By E15.5, interkinetic nuclear migration during the cell cycle was disrupted and the length of the cell cycle was significantly longer than normal in the Pax6Sey/Sey cortex, with a lengthening of S phase. Together, these results show that Pax6 is required in developing cortical progenitors to control the cell-cycle duration, the rate of progression from symmetrical to asymmetrical division and the onset of expression of neural-specific markers.

Microtubules and root protophloem ontogeny in wheat

Protophloem ontogeny in roots of Triticum aestivum has been investigated ultrastructurally. Each protophloem pole consists of three cells, a protophloem sieve element and two companion cells, all originating from a single precursor cell usually having a pentahedral shape. This protophloem mother cell (PMC) undergoes two successive asymmetrical divisions: the first one gives rise to a smaller cell that will differentiate into a companion cell, and a larger one that divides again asymmetrically yielding another companion cell and a protophloem sieve element. The latter divides once more, but now symmetrically, increasing the number of cells. Both asymmetrical and symmetrical divisions are preceded by preprophase microtubule bands (PMBs), well demarcated by a great number (more than 100 profiles in a single band section) of microtubules (MTs). The plane of a PMB coincides with that of the succeeding cell plate, which fuses with parent walls at sites previously occupied by the PMB. The strict correspondence between PMB and cell plate suggests that a cytokinesis the latter bisects the PMB cortical zone. The possible role of PMB cortical zone in positioning the cell plate and guiding its expanding edges towards predetermined sites is discussed in relation to recent discoveries in other anatomical situations. The plane of PMBs (and hence of divisions) changes from one division to the next, so that the three successive divisions occur in three spatial planes transversely to each other. This change is probably influenced by cell polarity. Prior to each asymmetrical division peri-nuclear MTs were observed besides the MTs of the PMB. They appear before the PMB organization and persist throughout preprophase, but they change their position and orientation in response to the transition from PMB to the spindle organization.

Organelle distribution and cell differentiation in the formation of stomatal complexes in barley

Interference – phase contrast observations of leaf epidermis in barley revealed the arrangement of organelles in cells that form stomatal complexes. Organelle movement and alignment were highly correlated with the process of cell division and differentiation that leads to mature stomata. Organelle rearrangement was most marked at the asymmetrical divisions that formed subsidiary cells of stomatal complexes.