Arabidopsis Reversibly Glycosylated Polypeptides 1 and 2 Are Essential for Pollen Development

Drakakaki, Georgia; Zabotina, Olga A.; Delgado, Ivan J.; Robert, Stéphanie; Keegstra, Kenneth; Raikhel, Natasha V.

doi:10.1104/pp.106.086363

Cited by 110 publications

(118 citation statements)

References 54 publications

Supporting

Mentioning

105

Contrasting

Unclassified

Order By: Relevance

“…Golgi/TGN vesicles were isolated by gently grinding leaves in chilled vesicle isolation buffer (HEPES 50 mM, pH 7.5, 0.45 M sucrose, 5 mM MgCl2, 1 mM DTT, 0.5% PVP (P2307, Sigma) and protease inhibitors (Roche)) and by separating the Golgi/TGN as previously described [65].…”

Section: Plant Materialsmentioning

confidence: 99%

Isolation and proteomic analysis of the SYP61 compartment reveal its role in exocytic trafficking in Arabidopsis

et al. 2011

Self Cite

View full text Add to dashboard Cite

The endomembrane system is a complex and dynamic intracellular trafficking network. It is very challenging to track individual vesicles and their cargos in real time; however, affinity purification allows vesicles to be isolated in their natural state so that their constituent proteins can be identified. Pioneering this approach in plants, we isolated the SYP61 trans-Golgi network compartment and carried out a comprehensive proteomic analysis of its contents with only minimal interference from other organelles. The proteome of SYP61 revealed the association of proteins of unknown function that have previously not been ascribed to this compartment. We identified a complete SYP61 SNARE complex, including regulatory proteins and validated the proteome data by showing that several of these proteins associated with SYP61 in planta. We further identified the SYP121-complex and cellulose synthases, suggesting that SYP61 plays a role in the exocytic trafficking and the transport of cell wall components to the plasma membrane. The presence of proteins of unknown function in the SYP61 proteome including ECHIDNA offers the opportunity to identify novel trafficking components and cargos. The affinity purification of plant vesicles in their natural state provides a basis for further analysis and dissection of complex endomembrane networks. The approach is widely applicable and can afford the study of several vesicle populations in plants, which can be compared with the SYP61 vesicle proteome.

show abstract

Section: Plant Materialsmentioning

confidence: 99%

Isolation and proteomic analysis of the SYP61 compartment reveal its role in exocytic trafficking in Arabidopsis

et al. 2011

Self Cite

View full text Add to dashboard Cite

show abstract

“…Tian et al (2004) present proof of principle for marker proteins targeted to numerous organelles, including MFP2, which was directed to peroxisomes. A number of studies have specifically used FTFLP to preserve the N-terminal targeting signal and the C-terminal retention motifs (Chen et al, 2005;Li et al, 2005;Drakakaki et al, 2006;Anand et al, 2007;Goh et al, 2007;Blakeslee et al, 2008;Gallavotti et al, 2008). We adapted the method for high-throughput analysis of full-length proteins via a transient expression assay.…”

Section: Gfp Localization Studiesmentioning

confidence: 99%

Novel Proteins, Putative Membrane Transporters, and an Integrated Metabolic Network Are Revealed by Quantitative Proteomic Analysis of Arabidopsis Cell Culture Peroxisomes

et al. 2008

View full text Add to dashboard Cite

Peroxisomes play key roles in energy metabolism, cell signaling, and plant development. A better understanding of these important functions will be achieved with a more complete definition of the peroxisome proteome. The isolation of peroxisomes and their separation from mitochondria and other major membrane systems have been significant challenges in the Arabidopsis (Arabidopsis thaliana) model system. In this study, we present new data on the Arabidopsis peroxisome proteome obtained using two new technical advances that have not previously been applied to studies of plant peroxisomes. First, we followed density gradient centrifugation with free-flow electrophoresis to improve the separation of peroxisomes from mitochondria. Second, we used quantitative proteomics to identify proteins enriched in the peroxisome fractions relative to mitochondrial fractions. We provide evidence for peroxisomal localization of 89 proteins, 36 of which have not previously been identified in other analyses of Arabidopsis peroxisomes. Chimeric green fluorescent protein constructs of 35 proteins have been used to confirm their localization in peroxisomes or to identify endoplasmic reticulum contaminants. The distribution of many of these peroxisomal proteins between soluble, membrane-associated, and integral membrane locations has also been determined. This core peroxisomal proteome from nonphotosynthetic cultured cells contains a proportion of proteins that cannot be predicted to be peroxisomal due to the lack of recognizable peroxisomal targeting sequence 1 (PTS1) or PTS2 signals. Proteins identified are likely to be components in peroxisome biogenesis, b-oxidation for fatty acid degradation and hormone biosynthesis, photorespiration, and metabolite transport. A considerable number of the proteins found in peroxisomes have no known function, and potential roles of these proteins in peroxisomal metabolism are discussed. This is aided by a metabolic network analysis that reveals a tight integration of functions and highlights specific metabolite nodes that most probably represent entry and exit metabolites that could require transport across the peroxisomal membrane.Within the plant cell, energy metabolism is mainly distributed among three distinct organelles: plastids, mitochondria, and peroxisomes. Although the proteomes of both plastids and mitochondria have been investigated extensively, comparatively little systematic analysis of the protein content of plant peroxisomes has been undertaken. The main obstacle for proteomics of plant peroxisomes is the availability of purified organelles from model plants that are also amenable to mass spectrometry (MS)-based identification by matching to protein sequence data. Whereas the preparation of peroxisomes in sufficient amounts and purity from spinach (Spinacia oleracea), cucumber (Cucumis sativus), pea (Pisum sativum), and soybean (Glycine max) for proteomic purposes is possible (Schwitzguebel and Siegenthaler, 1984;Corpas et al., 1994;Lopez-Huertas et al., 1999;Arai et al., 2008), the ...

show abstract

“…RGPs are likely delivered to the PD at the cell wall via the Golgi apparatus since pea and Arabidopsis RGPs 12,14,15 localize to the Golgi compartment, and disruption of Golgi function reduces accumulation of AtRGPs at PD in tobacco. 12 Overexpression of AtRGP2-GFP in stable tobacco transgenic plants causes stunting and chlorosis of source leaves due to increased starch accumulation and reduced chlorophyll content.…”

mentioning

confidence: 99%

“…14 To clarify the role of C1 RGPs in PD-mediated intercellular transport, we examined the function of Nicotiana benthamiana RGPs in their native context following virus-induced gene silencing (VIGS). AtRGP1 and AtRGP2 are very similar with 93% amino acid identity, suggesting they are redundant.…”

mentioning

confidence: 99%

Reduced levels of class 1 reversibly glycosylated polypeptide increase intercellular transport via plasmodesmata

Burch‐Smith

Cui

Zambryski

2012

Plant Signaling & Behavior

View full text Add to dashboard Cite

Maize and Arabidopsis thaliana class 1 reversibly glycosylated polypeptides ( C1 RGPs) are plasmodesmata-associated proteins. Previously, overexpression of Arabidopsis C1 RGP AtRGP2 in Nicotiana tabacum was shown to reduce intercellular transport of photoassimilate, resulting in stunted, chlorotic plants, and inhibition of local cell-to-cell spread of tobacco mosaic virus (TMV). Here, we used virus induced gene silencing to examine the effects of reduced levels of Plasmodesmata Structure and FunctionPlasmodesmata (PD) are membrane-lined cytoplasmic channels that connect contiguous plants cells, allowing transport of water, solutes, ions, nucleic acids and proteins between cells (for review see refs. 1-2). The importance of plasmodesmata to plant growth and development is demonstrated by the early lethality of mutants with altered PD structure and function.3,4 Electron microscopy reveals that PD consist largely of membranes and spaces (for review see ref. 5). Plasma membrane spans the cell wall between adjacent cells to form the outer limits of a given PD channel, and the ER of the adjacent two cells is continuous through the axial center of the channel, forming a specialized structure called the desmotubule (DT). Transport through PD is thought to predominantly occur in the space between the plasma membrane and DT, called the cytoplasmic sleeve, 6 while plant viruses also move along the membranes of the DT 7 and small molecules (~3 kDa) may move cell-to-cell via the lumen of the DT.

show abstract

Arabidopsis Reversibly Glycosylated Polypeptides 1 and 2 Are Essential for Pollen Development

Cited by 110 publications

References 54 publications

Isolation and proteomic analysis of the SYP61 compartment reveal its role in exocytic trafficking in Arabidopsis

Isolation and proteomic analysis of the SYP61 compartment reveal its role in exocytic trafficking in Arabidopsis

Novel Proteins, Putative Membrane Transporters, and an Integrated Metabolic Network Are Revealed by Quantitative Proteomic Analysis of Arabidopsis Cell Culture Peroxisomes

Reduced levels of class 1 reversibly glycosylated polypeptide increase intercellular transport via plasmodesmata

Contact Info

Product

Resources

About