scholarly journals The receptor-like kinases BAM1 and BAM2 promote the cell-to-cell movement of miRNA in the root stele to regulate xylem patterning

2019 ◽  
Author(s):  
Pengfei Fan ◽  
Hua Wang ◽  
Hao Xue ◽  
Tabata Rosas-Diaz ◽  
Weihua Tang ◽  
...  
Keyword(s):  
Cell Movement ◽  
Intercellular Movement ◽  
The Creation ◽  
Cell Spread ◽  
Root Stele

ABSTRACTXylem patterning in the root is established through the creation of opposing gradients of miRNAs and their targets, enabled by the cell-to-cell spread of the former. The miRNAs involved in xylem patterning, miR165/6, move through plasmodesmata, but how their trafficking is regulated remains elusive. Here, we describe that the receptor-like kinases BAM1/2 are required for the intercellular movement of miR165/6 in the stele and hence proper xylem patterning in the root.

scholarly journals Conversion in the Requirement of Coat Protein in Cell-to-Cell Movement Mediated by the Cucumber Mosaic Virus Movement Protein

2001 ◽  
Vol 75 (17) ◽  
pp. 8045-8053 ◽  
Author(s):  
Hideaki Nagano ◽  
Kazuyuki Mise ◽  
Iwao Furusawa ◽  
Tetsuro Okuno
Keyword(s):  
Amino Acids ◽  
Coat Protein ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Movement Protein ◽  
Cell Movement ◽  
Plant Viruses ◽  
Brome Mosaic Virus ◽  
Viral Movement ◽  
Intercellular Movement

ABSTRACT Plant viruses have movement protein (MP) gene(s) essential for cell-to-cell movement in hosts. Cucumber mosaic virus (CMV) requires its own coat protein (CP) in addition to the MP for intercellular movement. Our present results using variants of both CMV and a chimeric Brome mosaic virus with the CMV MP gene revealed that CMV MP truncated in its C-terminal 33 amino acids has the ability to mediate viral movement independently of CP. Coexpression of the intact and truncated CMV MPs extremely reduced movement of the chimeric viruses, suggesting that these heterogeneous CMV MPs function antagonistically. Sequential deletion analyses of the CMV MP revealed that the dispensability of CP occurred when the C-terminal deletion ranged between 31 and 36 amino acids and that shorter deletion impaired the ability of the MP to promote viral movement. This is the first report that a region of MP determines the requirement of CP in cell-to-cell movement of a plant virus.

scholarly journals The receptor-like kinases BAM1 and BAM2 are required for root xylem patterning

2021 ◽  
Vol 118 (12) ◽  
pp. e2022547118
Author(s):  
Pengfei Fan ◽  
Emmanuel Aguilar ◽  
Mariem Bradai ◽  
Hao Xue ◽  
Hua Wang ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Mirna Targets ◽  
The Creation ◽  
Cell Spread

Xylem patterning in the root is established through the creation of opposing gradients of miRNAs and their targets, transcripts of the HD-ZIP III family of transcriptions factors, enabled by the cell-to-cell spread of the former. The miRNAs regulating xylem patterning, miR165/6, move through plasmodesmata, but how their trafficking is regulated remains elusive. Here, we describe that simultaneous mutation of the plasma membrane- and plasmodesmata-localized receptor-like kinases (RLKs) BARELY ANY MERISTEM (BAM) 1 and 2 or expression of the geminivirus-encoded BAM1/2-interactor C4 results in higher accumulation and broader distribution of the HD-ZIP III transcripts despite normal total accumulation of miR165/6, and ultimately causes defects in xylem patterning, which depend on the function of the aforementioned miRNA targets. Taken together, our results show that BAM1 and BAM2 are redundantly required for proper xylem patterning in the Arabidopsis root, by ensuring the proper distribution and accumulation of miR165/6-targeted transcripts.

scholarly journals A plant receptor-like kinase promotes cell-to-cell spread of RNAi and is targeted by a virus

2017 ◽  
Author(s):  
Tabata Rosas-Diaz ◽  
Dan Zhang ◽  
Pengfei Fan ◽  
Liping Wang ◽  
Xue Ding ◽  
...  
Keyword(s):  
Cell Movement ◽  
Plant Cells ◽  
Tomato Yellow Leaf ◽  
Yellow Leaf ◽  
Systemic Spread ◽  
Receptor Like Kinase ◽  
Intercellular Spread ◽  
Leaf Curl Virus ◽  
Redundant Role ◽  
Cell Spread

ABSTRACTRNA interference (RNAi) in plants can move from cell to cell, allowing for systemic spread of an anti-viral immune response. How this cell-to-cell spread of silencing is regulated is currently unknown. Here, we describe that the C4 protein from Tomato yellow leaf curl virus has the ability to inhibit the intercellular spread of RNAi. Using this viral protein as a probe, we have identified the receptor-like kinase (RLK) BARELY ANY MERISTEM 1 (BAM1) as a positive regulator of the cell-to-cell movement of RNAi, and determined that BAM1 and its closest homologue, BAM2, play a redundant role in this process. C4 interacts with the intracellular domain of BAM1 and BAM2 at the plasma membrane and plasmodesmata, the cytoplasmic connections between plant cells, interfering with the function of these RLKs in the cell-to-cell spread of RNAi. Our results identify BAM1 as an element required for the cell-to-cell spread of RNAi and highlight that signalling components have been co-opted to play multiple functions in plants.

scholarly journals Identification of a Movement Protein of Rice Yellow Stunt Rhabdovirus

2005 ◽  
Vol 79 (4) ◽  
pp. 2108-2114 ◽  
Author(s):  
Yan-Wei Huang ◽  
Yun-Feng Geng ◽  
Xiao-Bao Ying ◽  
Xiao-Ying Chen ◽  
Rong-Xiang Fang
Keyword(s):  
Movement Protein ◽  
Specific Interaction ◽  
Protein Structures ◽  
Cell Movement ◽  
Protein Product ◽  
N Protein ◽  
Intercellular Movement ◽  
Main Component ◽  
Plant Rhabdovirus

ABSTRACT Rice yellow stunt rhabdovirus (RYSV) encodes seven genes in its negative-sense RNA genome in the order 3′-N-P-3-M-G-6-L-5′. The existence of gene 3 in the RYSV genome and an analogous gene(s) of other plant rhabdoviruses positioned between the P and M genes constitutes a unique feature for plant rhabdoviruses that is distinct from animal-infecting rhabdoviruses in which the P and M genes are directly linked. However, little is known about the function of these extra plant rhabdovirus genes. Here we provide evidence showing that the protein product encoded by gene 3 of RYSV, P3, possesses several properties related to a viral cell-to-cell movement protein (MP). Analyses of the primary and secondary protein structures suggested that RYSV P3 is a member of the “30K” superfamily of viral MPs. Biolistic bombardment transcomplementation experiments demonstrated that RYSV P3 can support the intercellular movement of a movement-deficient potexvirus mutant in Nicotiana benthamiana leaves. In addition, Northwestern blot analysis indicated that the RYSV P3 protein can bind single-stranded RNA in vitro, a common feature of viral MPs. Finally, glutathione S- transferase pull-down assays revealed a specific interaction between the RYSV P3 protein and the N protein which is a main component of the ribonucleocapsid, a subviral structure believed to be involved in the intercellular movement of plant rhabdoviruses. Together, these data suggest that RYSV P3 is likely a MP of RYSV, thus representing the first example of characterized MPs for plant rhabdoviruses.

scholarly journals Rice Dwarf Phytoreovirus Segment S6-Encoded Nonstructural Protein Has a Cell-to-Cell Movement Function

2004 ◽  
Vol 78 (10) ◽  
pp. 5382-5389 ◽  
Author(s):  
Yi Li ◽  
Yi M. Bao ◽  
Chun H. Wei ◽  
Zhen S. Kang ◽  
Yong W. Zhong ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Nonstructural Protein ◽  
Infectious Clone ◽  
Microprojectile Bombardment ◽  
Gene Segment ◽  
Cell Movement ◽  
Potato Virus X ◽  
Protein Product ◽  
Start Codon ◽  
Intercellular Movement

ABSTRACT Rice dwarf virus (RDV) is a member of the genus Phytoreovirus, which is composed of viruses with segmented double-stranded RNA genomes. Proteins that support the intercellular movement of these viruses in the host have not been identified. Microprojectile bombardment was used to determine which open reading frames (ORFs) support intercellular movement of a heterologous virus. A plasmid containing an infectious clone of Potato virus X (PVX) defective in cell-to-cell movement and expressing either β-glucuronidase or green fluorescent protein (GFP) was used for cobombardment with plasmids containing ORFs from RDV gene segments S1 through S12 onto leaves of Nicotiana benthamiana. Cell-to-cell movement of the movement-defective PVX was restored by cobombardment with a plasmid containing S6. In the absence of S6, no other gene segment supported movement. Identical results were obtained with Nicotiana tabacum, a host that allows fewer viruses to infect and spread within its tissue. S6 supported the cell-to-cell movement of the movement-defective PVX in sink and source leaves of N. benthamiana. A mutant S6 lacking the translation start codon did not complement the cell-to-cell movement of the movement-defective PVX. An S6 protein product (Pns6)-enhanced GFP fusion was observed near or within cell walls of epidermal cells from N. tabacum. By immunocytochemistry, unfused Pns6 was localized to plasmodesmata in rice leaves infected with RDV. S6 thus encodes a protein with characteristics identical to those of other viral proteins required for the cell-to-cell movement of their genome and therefore is likely required for the cell-to-cell movement of RDV.

scholarly journals A virus-targeted plant receptor-like kinase promotes cell-to-cell spread of RNAi

2018 ◽  
Vol 115 (6) ◽  
pp. 1388-1393 ◽  
Author(s):  
Tabata Rosas-Diaz ◽  
Dan Zhang ◽  
Pengfei Fan ◽  
Liping Wang ◽  
Xue Ding ◽  
...  
Keyword(s):  
Cell Movement ◽  
Tomato Yellow Leaf ◽  
Antiviral Immune Response ◽  
Systemic Spread ◽  
Receptor Like Kinase ◽  
Intercellular Spread ◽  
Leaf Curl Virus ◽  
Signaling Components ◽  
Redundant Role ◽  
Cell Spread

RNA interference (RNAi) in plants can move from cell to cell, allowing for systemic spread of an antiviral immune response. How this cell-to-cell spread of silencing is regulated is currently unknown. Here, we describe that the C4 protein from Tomato yellow leaf curl virus can inhibit the intercellular spread of RNAi. Using this viral protein as a probe, we have identified the receptor-like kinase (RLK) BARELY ANY MERISTEM 1 (BAM1) as a positive regulator of the cell-to-cell movement of RNAi, and determined that BAM1 and its closest homolog, BAM2, play a redundant role in this process. C4 interacts with the intracellular domain of BAM1 and BAM2 at the plasma membrane and plasmodesmata, the cytoplasmic connections between plant cells, interfering with the function of these RLKs in the cell-to-cell spread of RNAi. Our results identify BAM1 as an element required for the cell-to-cell spread of RNAi and highlight that signaling components have been coopted to play multiple functions in plants.

scholarly journals Development of Rice Stripe Tenuivirus Minireplicon Reverse Genetics Systems Suitable for Analyses of Viral Replication and Intercellular Movement

2021 ◽  
Vol 12 ◽  
Author(s):  
Xiaoyan Zhang ◽  
Kai Sun ◽  
Yan Liang ◽  
Shuo Wang ◽  
Kaili Wu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reporter Gene ◽  
Reverse Genetics ◽  
Ectopic Expression ◽  
Cell Movement ◽  
Rice Stripe Virus ◽  
Reporter Gene Expression ◽  
N Protein ◽  
Intercellular Movement ◽  
Negative Sense

Rice stripe virus (RSV), a tenuivirus with four negative-sense/ambisense genome segments, is one of the most devastating viral pathogens affecting rice production in many Asian countries. Despite extensive research, our understanding of RSV infection cycles and pathogenesis has been severely impaired by the lack of reverse genetics tools. In this study, we have engineered RSV minireplicon (MR)/minigenome cassettes with reporter genes substituted for the viral open reading frames in the negative-sense RNA1 or the ambisense RNA2-4 segments. After delivery to Nicotiana benthamiana leaves via agroinfiltration, MR reporter gene expression was detected only when the codon-optimized large viral RNA polymerase protein (L) was coexpressed with the nucleocapsid (N) protein. MR activity was also critically dependent on the coexpressed viral suppressors of RNA silencing, but ectopic expression of the RSV-encoded NS3 silencing suppressor drastically decreased reporter gene expression. We also developed intercellular movement-competent MR systems with the movement protein expressed either in cis from an RNA4-based MR or in trans from a binary plasmid. Finally, we generated multicomponent replicon systems by expressing the N and L proteins directly from complementary-sense RNA1 and RNA3 derivatives, which enhanced reporter gene expression, permitted autonomous replication and intercellular movement, and reduced the number of plasmids required for delivery. In summary, this work enables reverse genetics analyses of RSV replication, transcription, and cell-to-cell movement and provides a platform for engineering more complex recombinant systems.

scholarly journals P3N-PIPO Interacts with P3 via the Shared N-Terminal Domain To Recruit Viral Replication Vesicles for Cell-to-Cell Movement

2020 ◽  
Vol 94 (8) ◽  
Author(s):  
Mengzhu Chai ◽  
Xiaoyun Wu ◽  
Jiahui Liu ◽  
Yue Fang ◽  
Yameng Luan ◽  
...  
Keyword(s):  
Viral Replication ◽  
Mosaic Virus ◽  
Essential Role ◽  
Movement Protein ◽  
Cell Movement ◽  
Plant Viruses ◽  
Turnip Mosaic Virus ◽  
Limited Information ◽  
Intercellular Movement ◽  
Model Virus

ABSTRACT P3N-PIPO, the only dedicated movement protein (MP) of potyviruses, directs cylindrical inclusion (CI) protein from the cytoplasm to the plasmodesma (PD), where CI forms conical structures for intercellular movement. To better understand potyviral cell-to-cell movement, we further characterized P3N-PIPO using Turnip mosaic virus (TuMV) as a model virus. We found that P3N-PIPO interacts with P3 via the shared P3N domain and that TuMV mutants lacking the P3N domain of either P3N-PIPO or P3 are defective in cell-to-cell movement. Moreover, we found that the PIPO domain of P3N-PIPO is sufficient to direct CI to the PD, whereas the P3N domain is necessary for localization of P3N-PIPO to 6K2-labeled vesicles or aggregates. Finally, we discovered that the interaction between P3 and P3N-PIPO is essential for the recruitment of CI to cytoplasmic 6K2-containing structures and the association of 6K2-containing structures with PD-located CI inclusions. These data suggest that both P3N and PIPO domains are indispensable for potyviral cell-to-cell movement and that the 6K2 vesicles in proximity to PDs resulting from multipartite interactions among 6K2, P3, P3N-PIPO, and CI may also play an essential role in this process. IMPORTANCE Potyviruses include numerous economically important viruses that represent approximately 30% of known plant viruses. However, there is still limited information about the mechanism of potyviral cell-to-cell movement. Here, we show that P3N-PIPO interacts with and recruits CI to the PD via the PIPO domain and interacts with P3 via the shared P3N domain. We further report that the interaction of P3N-PIPO and P3 is associated with 6K2 vesicles and brings the 6K2 vesicles into proximity with PD-located CI structures. These results support the notion that the replication and cell-to-cell movement of potyviruses are processes coupled by anchoring viral replication complexes at the entrance of PDs, which greatly increase our knowledge of the intercellular movement of potyviruses.

scholarly journals A bootstrap approach is a superior statistical method for the comparison of cell-to-cell movement data

2020 ◽  
Author(s):  
Matthew G. Johnston ◽  
Christine Faulkner
Keyword(s):  
False Positive ◽  
Bootstrap Method ◽  
False Positive Rate ◽  
Cell Movement ◽  
Wilcoxon Test ◽  
Data Sets ◽  
Movement Data ◽  
Positive Rate ◽  
Intercellular Movement ◽  
The Bootstrap Method

SummaryPlasmodesmata are an increasing focus of plant research, and plant physiologists frequently aim to understand the dynamics of intercellular movement and plasmodesmal function. For this, experiments that measure the spread of GFP between cells are commonly performed to indicate whether plasmodesmata are more open or closed in different conditions or in different genotypes.We propose cell-to-cell movement data sets are better analysed by a bootstrap method that tests the null hypothesis that means (or medians) are the same between two conditions, instead of the commonly used Mann-Whitney-Wilcoxon test. We found that that with hypothetical distributions similar to cell-to-cell movement data, the Mann-Whitney-Wilcoxon produces a false positive rate of 17% while the bootstrap method maintains a false positive at the set rate of 5% under the same circumstances. Here we present this finding, as well as our rationale, an explanation of the bootstrap method and an R script for easy use. We have further demonstrated its use on published datasets from independent laboratories.

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