An official website of the United States government
Abstract Numerous cell surface receptors and receptor-like proteins (RLPs) undergo activation or deactivation via a transmembrane domain (TMD). A subset of plant RLPs distinctively localizes to the plasma membrane-lined pores called plasmodesmata. Those RLPs include theArabidopsis thalianaPlasmodesmata-located protein (PDLP) 5, which is well known for its vital function regulating plasmodesmal gating and molecular movement between cells. In this study, we report that the TMD, although not a determining factor for the plasmodesmal targeting, serves essential roles for the PDLP5 function. In addition to its role for membrane anchoring, the TMD mediates PDLP5 self-interaction and carries an evolutionarily conserved motif that is essential for PDLP5 to regulate cell-to-cell movement. Computational modeling-based analyses suggest that PDLP TMDs have high propensities to dimerize. We discuss how a specific mode(s) of TMD dimerization might serve as a common mechanism for PDLP5 and other PDLP members to regulate cell-to-cell movement.
more »
« less
Targeting of plasmodesmal proteins requires unconventional signals
Abstract Effective cellular signaling relies on precise spatial localization and dynamic interactions among proteins in specific subcellular compartments or niches, such as cell-to-cell contact sites and junctions. In plants, endogenous and pathogenic proteins gained the ability to target plasmodesmata, membrane-lined cytoplasmic connections, through evolution to regulate or exploit cellular signaling across cell wall boundaries. For example, the receptor-like membrane protein PLASMODESMATA-LOCATED PROTEIN 5 (PDLP5), a potent regulator of plasmodesmal permeability, generates feed-forward or feed-back signals important for plant immunity and root development. However, the molecular features that determine the plasmodesmal association of PDLP5 or other proteins remain largely unknown, and no protein motifs have been identified as plasmodesmal targeting signals. Here, we developed an approach combining custom-built machine-learning algorithms and targeted mutagenesis to examine PDLP5 in Arabidopsis thaliana and Nicotiana benthamiana. We report that PDLP5 and its closely related proteins carry unconventional targeting signals consisting of short stretches of amino acids. PDLP5 contains 2 divergent, tandemly arranged signals, either of which is sufficient for localization and biological function in regulating viral movement through plasmodesmata. Notably, plasmodesmal targeting signals exhibit little sequence conservation but are located similarly proximal to the membrane. These features appear to be a common theme in plasmodesmal targeting.
more »
« less
- Award ID(s):
- 1820103
- PAR ID:
- 10437997
- Publisher / Repository:
- Oxford University Press
- Date Published:
- Journal Name:
- The Plant Cell
- Volume:
- 35
- Issue:
- 8
- ISSN:
- 1040-4651
- Page Range / eLocation ID:
- p. 3035-3052
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
null; Murali T., Narasimhan G.; Rajasekaran S., Skums P.; null (Ed.)Subcellular localization plays important roles in protein’s functioning. In this paper, we developed a hidden Markov model to detect de novo signals in protein sequences that target at a particular cellular location: plasmodesmata. We also developed a support vector machine to classify plasmodesmata located proteins (PDLPs) in Arabidopsis, and devised a decision-tree approach to combine the SVM and HMM for better classification performance. The methods achieved high performance with ROC score 0.99 in cross-validation test on a set of 360 type I transmembrane proteins in Arabidopsis. The predicted PD targeting signals in one PDLP have been experimentally verified.more » « less
-
Abstract Plasmodesmata connect adjoining plant cells, allowing molecules to move between the connected cells for communication and sharing resources. It has been well established that the plant polysaccharide callose is deposited at plasmodesmata, regulating their aperture and function. Among proteins involved in maintaining callose homeostasis, PLASMODESMATA-LOCATED PROTEINSs (PDLPs) promote callose deposition at plasmodesmata. This study explored the function of PDLP5 and PDLP6 in different cell types. We discovered that PDLP5 and PDLP6 are expressed in nonoverlapping cell types in Arabidopsis (Arabidopsis thaliana). The overexpression of PDLP5 and PDLP6 results in the overaccumulation of plasmodesmal callose at different cell interfaces, indicating that PDLP5 and PDLP6 are active in different cell types. We also observed 2 distinct patterns of starch accumulation in mature leaves of PDLP5 and PDLP6 overexpressors. An enzyme-catalyzed proximity labeling approach was used to identify putative functional partners of the PDLPs. We identified SUCROSE SYNTHASE 6 (SUS6) as a functional partner of PDLP6 in the vasculature. We further demonstrated that PDLP6 physically and genetically interacts with SUS6. In addition, CALLOSE SYNTHASE 7 (CALS7) physically interacts with SUS6 and PDLP6. Genetic interaction studies showed that CALS7 is required for PDLP6 function. We propose that PDLP6 functions with SUS6 and CALS7 in the vasculature to regulate plasmodesmal function.more » « less
-
Abstract Optogenetic tools are created to control RhoA GTPase, a central regulator of actin organization and actomyosin contractility. RhoA GTPase, or its upstream activator ARHGEF11, is fused to BcLOV4, a photoreceptor that can be dynamically recruited to the plasma membrane by a light‐regulated protein‐lipid electrostatic interaction with the inner leaflet. Direct membrane recruitment of these proteins induces potent contractile signaling sufficient to separate adherens junctions with as little as one pulse of blue light. Induced cytoskeletal morphology changes are dependent on the alignment of the spatially patterned stimulation with the underlying cell polarization. RhoA‐mediated cytoskeletal activation drives yes‐associated protein (YAP) nuclear localization within minutes and consequent mechanotransduction verified by YAP‐transcriptional enhanced associate domain transcriptional activity. These single‐transgene tools do not require protein binding partners for dynamic membrane localization and permit spatiotemporally precise control over RhoA signaling to advance the study of its diverse regulatory roles in cell migration, morphogenesis, and cell cycle maintenance.more » « less
-
null (Ed.)Plasmodesmata are membrane-lined cytoplasmic passageways that facilitate the movement of nutrients and various types of molecules between cells in the plant. They are highly dynamic channels, opening or closing in response to physiological and developmental stimuli or environmental challenges such as biotic and abiotic stresses. Accumulating evidence supports the idea that such dynamic controls occur through integrative cellular mechanisms. Currently, a few fluorescence-based methods are available that allow monitoring changes in molecular movement through plasmodesmata. In this chapter, following a brief introduction to those methods, we provide a detailed step-by-step protocol for the Drop-ANd-See (DANS) assay, which is advantageous when it is desirable to measure plasmodesmal permeability non-invasively, in situ and in real-time. We discuss the experimental conditions one should consider to produce reliable and reproducible DANS results along with troubleshooting ideas.more » « less
