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de Keijzer, J.

Publications and source records attributed to de Keijzer, J..

3 recordsLinked to original sources

The proximal proteome of FLOWERING LOCUS T LIKE 1 during rice panicle development suggests cell-to-cell mobility features

Flowering is promoted by perception of favorable environmental stimuli. In rice, exposure of leaves to short days induces the differentiation of a branched inflorescence called panicle at the shoot apical meristem (SAM). Systemic communication from the leaves to the SAM is mediated by members of the phosphatidylethanolamine-binding protein (PEBP) family, including HEADING DATE 3a (Hd3a) and RICE FLOWERING LOCUS T 1 (RFT1), commonly referred to as florigens. Florigens are [~]20kDa proteins translated in leaf companion cells and loaded into the phloematic stream, through which they reach the SAM. Once in meristematic cells, they are translocated to the nucleus, where they form higher-order protein complexes that include transcription factors and drive transcriptional reprogramming of meristematic cells. The activity of Hd3a and RFT1 at the SAM is partly mediated by FLOWERING LOCUS T LIKE 1 (FTL1) encoding a florigen-like protein required to accelerate flowering and establish the branching pattern of the panicle. The photoperiodic regulatory network is largely based on protein-protein interactions (PPIs), that take place in different tissues and in response to environmental variation. Capturing the full extent of these interactions for a specific protein and in vivo is a challenging task. Here, we developed a protocol of Proximity Labeling (PL) to study the interactome of FTL1 in developing panicles. We show that FTL1 associates with, or is proximal to, several proteins implicated in vesicular trafficking, microtubule binding and transcriptional regulation. Using a transient system, we demonstrate that FTL1 can actively move between cells, similarly to Hd3a and RFT1. This study establishes a protocol to implement PL for the study of a dynamic process during rice reproductive development, identifies interactors of FTL1, and suggests novel features possibly linked to its function.

developmental biology↗

Comparative phyloproteomics identifies conserved plasmodesmal proteins

Plasmodesmata connect neighbouring plant cells across the cell wall. They are cytosolic bridges, lined by the plasma membrane and traversed by endoplasmic reticulum to connect these cell components between cells and tissues. While plasmodesmata are notoriously difficult to extract, tissue fractionation and proteomic analyses have yielded valuable knowledge of their composition. Most proteomic profiles originate from cell suspension cultures in which simple plasmodesmata dominate and have been exclusively generated from dicotyledonous plant species. Here we have generated two novel proteomes to expand tissue and taxonomic representation of plasmodesmata: one from mature Arabidopsis leaves and one from the moss Physcomitrium patens. We have leveraged these and existing data to perform a comparative analysis that, owing to comparing proteomes from an expanded taxonomic tree, allowed us to identify conserved protein families that are associated with plasmodesmata that likely serve as core structural or functional components. Thus, we identified {beta}-1,3-glucanases, C2 lipid-binding proteins and tetraspanins as core plasmodesmal components, with proteins from P. patens and Arabidopsis maintaining plasmodesmal association across diverse species. Our approach has not only identified elements of a conserved, core plasmodesmal proteome, but also demonstrated the added power offered by comparative analysis. Conserved plasmodesmal proteins establish a basis upon which ancient plasmodesmal function can be further investigated to determine the essential roles these structures play in multicellular organism physiology in the green lineages.

plant biology↗

Exocyst subunit Sec6 is positioned by microtubule overlaps in the moss phragmoplast prior to the arrival of cell plate membrane

During plant cytokinesis a radially expanding membrane-enclosed cell plate is formed from fusing vesicles that compartmentalizes the cell in two. How fusion is spatially restricted to the site of cell plate formation is unknown. Aggregation of cell-plate membrane starts near regions of microtubule overlap within the bipolar phragmoplast apparatus of the moss Physcomitrella patens. Since vesicle fusion generally requires coordination of vesicle tethering and subsequent fusion activity we analysed the subcellular localization of several subunits of the exocyst, a tethering complex active during plant cytokinesis. We found that Sec6, but neither Sec3 or Sec5 subunits localized to microtubule overlap regions in advance of cell plate construction started in moss. Moreover, Sec6 exhibited a conserved physical interaction with an orthologue of the Sec1/Munc18 protein KEULE, an important regulator for cell-plate membrane vesicle fusion in Arabidopsis. Recruitment of PpKEULE and vesicles to the early cell plate was delayed upon Sec6 gene silencing. Our findings thus suggest that vesicle-vesicle fusion is in part enabled by a pool of exocyst subunits at microtubule overlaps that is recruited independent of the delivery of vesicles.\n\nSummary statementWe performed a time-resolved localization screen of multiple subunits of the exocyst complex throughout moss cytokinesis and show that each subunit has a unique spatiotemporal recruitment pattern.

cell biology↗