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Expression profiles of embryo-defective Arabidopsis thaliana mutants increased size exclusion limit1 (ise1) and ise2

ABSTRACT: We use Arabidopsis thaliana embryogenesis as a model system for studying intercellular transport via plasmodesmata (PD). A forward genetic screen for altered PD transport identified ise1 and ise2 mutants with increased intercellular transport of fluorescent 10-kDa tracers. Both ise1 and ise2 exhibit increased formation of twinned and branched PD. ISE1 encodes a mitochondrial DEAD-box RNA helicase, while ISE2 encodes a DEVH-type RNA helicase. Here we show that ISE2 foci are localized to the chloroplast stroma. Surprisingly, plastid development is defective in both ise1 and ise2 mutant embryos. In an effort to understand how RNA helicases that localize to different organelles have similar impacts on plastid and PD development/function we performed whole genome expression analyses. The most significantly affected class of transcripts in both mutants encodes products that target to and enable plastid function. These results reinforce the importance of plastid-mitochondria-nucleus crosstalk, add PD as a critical player in the plant cell communication network, and thereby illuminate a new signaling pathway, dubbed organelle-nucleus-plasmodesmata signaling (ONPS). Several genes with roles in cell wall synthesis and modification are also differentially expressed in both mutants, providing new targets for investigating PD development and function.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE33558 | GEO | 2011/11/09

SECONDARY ACCESSION(S): PRJNA148875

REPOSITORIES: GEO

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Expression profiles of embryo-defective Arabidopsis thaliana mutants increased size exclusion limit1 (ise1) and ise2

Project description:We use Arabidopsis thaliana embryogenesis as a model system for studying intercellular transport via plasmodesmata (PD). A forward genetic screen for altered PD transport identified ise1 and ise2 mutants with increased intercellular transport of fluorescent 10-kDa tracers. Both ise1 and ise2 exhibit increased formation of twinned and branched PD. ISE1 encodes a mitochondrial DEAD-box RNA helicase, while ISE2 encodes a DEVH-type RNA helicase. Here we show that ISE2 foci are localized to the chloroplast stroma. Surprisingly, plastid development is defective in both ise1 and ise2 mutant embryos. In an effort to understand how RNA helicases that localize to different organelles have similar impacts on plastid and PD development/function we performed whole genome expression analyses. The most significantly affected class of transcripts in both mutants encodes products that target to and enable plastid function. These results reinforce the importance of plastid-mitochondria-nucleus crosstalk, add PD as a critical player in the plant cell communication network, and thereby illuminate a new signaling pathway, dubbed organelle-nucleus-plasmodesmata signaling (ONPS). Several genes with roles in cell wall synthesis and modification are also differentially expressed in both mutants, providing new targets for investigating PD development and function. Three biological replicates each of either ise1 or ise2 mutant seeds vs. sister wild-type controls

2011-11-09 | E-GEOD-33558 | biostudies-arrayexpress

Assaying effector cell-to-cell mobility in plant tissues identifies hypermobility and indirect manipulation of plasmodesmata

Project description:In plants, plasmodesmata establish cytoplasmic continuity between cells to allow for communication and resource exchange across the cell wall. While plant pathogens use plasmodesmata as a pathway for both molecular and physical invasion, the benefits of molecular invasion (cell-to-cell movement of pathogen effectors) are poorly understood. To establish a methodology for identification and characterization of the cell-to-cell mobility of effectors, we performed a quantitative live imaging-based screen of candidate effectors of the fungal pathogen Colletotrichum higginsianum. We predicted C. higginsianum effectors by their expression profiles, the presence of a secretion signal, and their predicted and in planta localization when fused to GFP. We assayed for cell-to-cell mobility of nucleo-cytosolic effectors and identified 14 that are cell-to-cell mobile. We identified that 3 of these effectors are “hypermobile”, showing cell-to-cell mobility greater than expected for a protein of its size. To explore the mechanism of hypermobility we chose two hypermobile effectors and measured their impact on plasmodesmata function and found that even though they show no direct association with plasmodesmata, each increases the transport capacity of plasmodesmata. Thus, our methods for quantitative analysis of cell-to-cell mobility of candidate microbe-derived36effectors, or any suite of host proteins, can identify cell-to-cell hypermobility and offer greater understanding of how proteins affect plasmodesmal function and intercellular connectivity.

2023-11-27 | GSE248400 | GEO

Choline transporter-like1 is required for plasmodesmal targeting of Potato leafroll virus movement protein MP17 and long-distance assimilate transport

Project description:Macromolecular trafficking and cell-to-cell communication in plants occurs via plasmodesmata (PD). Currently, little is known about the proteins defining these sophisticated cell-to-cell contacts. To uncover proteins required for PD structure and function we made use of the 17 kDa movement protein (MP17) of the Potato leafroll virus (PLRV). The protein is required for cell-to-cell movement of the virus and localizes specifically to branched PD in source tissues. By forward genetic screening for Arabidopsis mutants with altered PD binding of MP17, several mutants were found. Map-based cloning of one of these mutants revealed a mutation in the choline transporter-like 1 (CHER1) protein. The mutation abolished plasma membrane localization of the protein. As consequence phosphatidylcholine levels and PD binding of MP17 decreased. Transcriptome analysis revealed a down-regulation of defense genes and genes involved in the synthesis of very long chain fatty acids, indicating an impairment of lipid signaling. Furthermore, cher1 mutants showed a reduced assimilate export and stunted growth. These findings highlight the emerging role of phospholipids, especially in the context of PD structure and function as well as potential binding sites for plasma membrane- and PD-associated proteins, which has been neglected for a long time. light exposed source leaves from wild type Col-0, T-DNA insertion line cher1-4 and mutant cher1-5 (10 weeks old). Samples: per pool 2 source leaves from 4 different plants, taken in the afternoon; wildtype (WT): Col-0, cher1-4: T-DNA insertion line SALK-065853 (ecotype Col-0, kanamycin resistance), cher1-5: Col-0 with GGA -->GAA at position 740 of cds

2016-06-01 | E-GEOD-61595 | biostudies-arrayexpress

Choline transporter-like1 is required for plasmodesmal targeting of Potato leafroll virus movement protein MP17 and long-distance assimilate transport

2016-06-01 | GSE61595 | GEO

Arabidopsis thaliana cultivar:Columbia-0

Project description:Plasmodesmata localizing proteins regulate transport and signaling during systemic immunity

| PRJNA315406 | ENA

Multiple C2 domains and Transmembrane region Proteins (MCTPs) tether membranes at plasmodesmata

Project description:In eukaryotes, membrane contact sites (MCS) allow direct communication between organelles. Plants have evolved unique MCS, the plasmodesmata intercellular pores, which combine organelle tethering with regulation of cell-to-cell signalling, the molecular mechanisms of which remains unknown. Here, we identify Multiple C2 domains and Transmembrane region Proteins (MCTPs) as tethers that link the endoplasmic reticulum (ER) to the plasma membrane (PM) within plasmodesmata. We report that MCTPs, including MCTP3 and 4 recently identified as modulators of SHOOTMERISTEMLESS trafficking, are ER-anchored proteins that cluster at plasmodesmata. MCTPs insert into the ER via their transmembrane region whilst their C2 domains dock to the PM through interaction with anionic phospholipids. A mctp3/4 loss-of function mutant induces plant developmental defects while MCTP4 expression in a yeast .tether mutant partially restores ER-PM tethering. Our data suggest that MCTPs are unique membrane tethers controlling both ER-PM contacts and cell-cell signalling.

2019-07-08 | PXD013999 | Pride

Proteomic analysis of plasmodesmata from Populus cell suspension cultures

Project description:In this project, we have investigated the proteome of the plasmodesmata (PD) enriched membrane fraction from poplar cell suspension cultures and quantitatively compared its protein composition with other membrane fractions. We build on the previously published literature on the PD proteome by using semiquantitative methods and, for the first time, demonstrate in vitro callose synthase activity from Populus PD proteins. To our knowledge, this study is the first proteomic investigation of the PD performed on a tree species.

2018-12-31 | PXD010692 | Pride

Sigma2 and Sigma6-mediated plastid retrograde signals control nuclear gene expression in Arabidopsis

Project description:Retrograde signaling from the chloroplast to the nucleus is necessary to regulate the chloroplast proteome during development and fluctuating environmental conditions. Although the specific chloroplast process(es) that must occur and the nature of the signal(s) that exits the chloroplast are not well understood, previous studies using drug inhibitors of chloroplast biogenesis have revealed that normal chloroplast development is required to express Photosynthesis Associated Nuclear Genes (PhANGs). In an attempt to determine which specific steps in chloroplast development are involved in retrograde signaling, we analyzed Arabidopsis mutants defective in the six genes encoding sigma factor (Sig) proteins that are utilized by the plastid-encoded RNA polymerase to transcribe specific sets of plastid genes. Here, we demonstrate that both Sig2 and Sig6 have partially redundant roles in not only plastid transcription, but also tetrapyrrole synthesis and retrograde signaling to control PhANG expression. Normal PhANG expression can be partly restored in the sig2 mutant by increasing heme synthesis. Furthermore, there is a genetic interaction between Sig and GUN (genomes uncoupled) genes to generate chloroplast-retrograde signals. These results demonstrate that defective plastid transcription is the source of at least two retrograde signals to the nucleus; one involving tetrapyrrole synthesis and the other involving the accumulation of an unknown plastid transcript. We also propose that the study of sig mutants (with defects in the expression of specific plastid genes) provides a new genetic system, which avoids the use of harsh inhibitors and their potential side effects, to monitor developmental retrograde signaling and to elucidate its mechanisms.

2012-09-18 | E-MTAB-4338 | biostudies-arrayexpress

Comparative proteomics of sink and source cell wall

Project description:Plants as multi cellular organisms are composed of various specialized cell types. Most of these cell types are symplasmically connected via plasmodesmata (PD) allowing exchange of nutrients and signal molecules. To date, the molecular composition of PD is not well understood. A comprehensive and comparative proteomic approach was followed to identify proteins providing new insights into the composition of these important regulatory structures.

2017-09-06 | PXD007282 | Pride

Comparative proteomics of plasmodesmata

2017-09-05 | PXD006880 | Pride

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