Differing requirements for actin and myosin by plant viruses for sustained intercellular movement - PubMed (original) (raw)
Differing requirements for actin and myosin by plant viruses for sustained intercellular movement
Phillip A Harries et al. Proc Natl Acad Sci U S A. 2009.
Abstract
The actin cytoskeleton has been implicated in the intra- and intercellular movement of a growing number of plant and animal viruses. However, the range of viruses influenced by actin for movement and the mechanism of this transport are poorly understood. Here we determine the importance of microfilaments and myosins for the sustained intercellular movement of a group of RNA-based plant viruses. We demonstrate that the intercellular movement of viruses from different genera [tobacco mosaic virus (TMV), potato virus X (PVX), tomato bushy stunt virus (TBSV)], is inhibited by disruption of microfilaments. Surprisingly, turnip vein-clearing virus (TVCV), a virus from the same genus as TMV, did not require intact microfilaments for normal spread. To investigate the molecular basis for this difference we compared the subcellular location of GFP fusions to the 126-kDa protein and the homologous 125-kDa protein from TMV and TVCV, respectively. The 126-kDa protein formed numerous large cytoplasmic inclusions associated with microfilaments, whereas the 125-kDa protein formed few small possible inclusions, none associated with microfilaments. The dependence of TMV, PVX, and TBSV on intact microfilaments for intercellular movement led us to investigate the role of myosin motors in this process. Virus-induced gene silencing of the Nicotiana benthamiana myosin XI-2 gene, but not three other myosins, inhibited only TMV movement. These results indicate that RNA viruses have evolved differently in their requirements for microfilaments and the associated myosin motors, in a manner not correlated with predicted phylogeny.
Conflict of interest statement
The authors declare no conflict of interest.
Figures
Fig. 1.
The effect of LatB on virus spread. (A) Representative images showing GFP lesions formed upon infection with the indicated viruses either in the absence (-) or presence (+) of 5 μM LatB. All images were taken at 6 dpi. (Scale bar, 1 mm.) (B–E) Lesion areas were quantified to determine the effect of LatB on the cell-to-cell movement of (B) TMV, (C) PVX, (D) TBSV, and (E) TVCV at 2, 4, and 6 dpi. N. benthamiana leaf tissue was infiltrated with either the actin inhibitor LatB (circles) or a DMSO buffer control (squares). Bars represent standard errors for 15 lesions per treatment.
Fig. 2.
The TVCV 125-kDa protein does not form numerous large inclusions like the TMV 126-kDa protein. Representative images showing TMV 126-kDa protein (A) and TVCV 125-kDa protein (B) GFP fusions expressed in N. benthamiana leaf epidermal cells 3 days following agrobacterium infiltration. The positions of TMV 126-kDa protein bodies (A) and potential TVCV 125-kDa protein bodies (B) are indicated with arrows. Red fluorescent bodies are chloroplasts. (Scale bar, 25 μm.)
Fig. 3.
VIGS of individual myosin genes. Quantitative RT-PCR was used to determine the relative expression ratio of target genes (myosin VIII-1, myosin VIII-2, myosin XI-2, and myosin XI-F) in lines treated with the indicated TRV silencing constructs versus a TRV control not expressing a myosin fragment. Elongation factor 1α served as an internal loading control for each sample. Expression analysis was performed on extracts from systemic leaves at 18 dpi with TRV constructs. Bars represent means ± standard errors for three replicates per treatment. Analysis of variance followed by an lsd calculation was used to determine significant differences between treatments. Different letters above the bars indicate significant differences (P = 0.05). The experiment was repeated at least once for each TRV silencing construct.
Fig. 4.
TMV utilizes a distinct myosin for virus spread in N. benthamiana. GFP lesion areas reflect the cell-to-cell movement of (A) TMV, (B) TVCV, (C) PVX, and (D) TBSV in N. benthamiana leaves silenced for individual myosin genes (VIII-1, VIII-2, XI-2, XI-F) via TRV VIGS. Plants inoculated with wild-type TRV (WT TRV) or buffer (Mock) were controls. The area of GFP fluorescent lesions (mm2) in inoculated leaves was determined at 3 dpi for tissues carrying a systemic infection with the TRV VIGS vector (approximately 18 dpi). Bars represent means ± standard errors for three replicates per treatment. Analysis of variance followed by an lsd calculation was used to determine significant differences between treatments. Different letters above the bars indicate significant differences (P = 0.05). The experiment was repeated at least once for each virus challenge.
Similar articles
- Inhibition of tobacco mosaic virus movement by expression of an actin-binding protein.
Hofmann C, Niehl A, Sambade A, Steinmetz A, Heinlein M. Hofmann C, et al. Plant Physiol. 2009 Apr;149(4):1810-23. doi: 10.1104/pp.108.133827. Epub 2009 Feb 13. Plant Physiol. 2009. PMID: 19218363 Free PMC article. - The tobamovirus Turnip Vein Clearing Virus 30-kilodalton movement protein localizes to novel nuclear filaments to enhance virus infection.
Levy A, Zheng JY, Lazarowitz SG. Levy A, et al. J Virol. 2013 Jun;87(11):6428-40. doi: 10.1128/JVI.03390-12. Epub 2013 Mar 27. J Virol. 2013. PMID: 23536678 Free PMC article. - Turnip vein clearing virus movement protein nuclear activity: Do Tobamovirus movement proteins play a role in immune response suppression?
Levy A. Levy A. Plant Signal Behav. 2015;10(10):e1066951. doi: 10.1080/15592324.2015.1066951. Epub 2015 Aug 3. Plant Signal Behav. 2015. PMID: 26237173 Free PMC article. - Actin microfilament dynamics and actin side-binding proteins in plants.
Higaki T, Sano T, Hasezawa S. Higaki T, et al. Curr Opin Plant Biol. 2007 Dec;10(6):549-56. doi: 10.1016/j.pbi.2007.08.012. Epub 2007 Nov 1. Curr Opin Plant Biol. 2007. PMID: 17936064 Review. - Actin-myosin XI: an intracellular control network in plants.
Duan Z, Tominaga M. Duan Z, et al. Biochem Biophys Res Commun. 2018 Nov 25;506(2):403-408. doi: 10.1016/j.bbrc.2017.12.169. Epub 2018 Jan 5. Biochem Biophys Res Commun. 2018. PMID: 29307817 Review.
Cited by
- Viroid Replication, Movement, and the Host Factors Involved.
Zhang Y, Nie Y, Wang L, Wu J. Zhang Y, et al. Microorganisms. 2024 Mar 12;12(3):565. doi: 10.3390/microorganisms12030565. Microorganisms. 2024. PMID: 38543616 Free PMC article. Review. - Advances in Understanding the Mechanism of Cap-Independent Cucurbit Aphid-Borne Yellows Virus Protein Synthesis.
Truniger V, Pechar GS, Aranda MA. Truniger V, et al. Int J Mol Sci. 2023 Dec 18;24(24):17598. doi: 10.3390/ijms242417598. Int J Mol Sci. 2023. PMID: 38139425 Free PMC article. - A sword or a buffet: plant endomembrane system in viral infections.
Jovanović I, Frantová N, Zouhar J. Jovanović I, et al. Front Plant Sci. 2023 Aug 11;14:1226498. doi: 10.3389/fpls.2023.1226498. eCollection 2023. Front Plant Sci. 2023. PMID: 37636115 Free PMC article. Review. - Manipulation of the Cellular Membrane-Cytoskeleton Network for RNA Virus Replication and Movement in Plants.
He R, Li Y, Bernards MA, Wang A. He R, et al. Viruses. 2023 Mar 14;15(3):744. doi: 10.3390/v15030744. Viruses. 2023. PMID: 36992453 Free PMC article. Review. - Distinct Mechanisms of Endomembrane Reorganization Determine Dissimilar Transport Pathways in Plant RNA Viruses.
Solovyev AG, Atabekova AK, Lezzhov AA, Solovieva AD, Chergintsev DA, Morozov SY. Solovyev AG, et al. Plants (Basel). 2022 Sep 15;11(18):2403. doi: 10.3390/plants11182403. Plants (Basel). 2022. PMID: 36145804 Free PMC article. Review.
References
- Verchot-Lubicz J, Ye CM, Bamunusinghe D. Molecular biology of potexviruses: Recent advances. J Gen Virol. 2007;88:1643–1655. - PubMed
- Hofmann C, Sambade A, Heinlein M. Plasmodesmata and intercellular transport of viral RNA. Biochem Soc Trans. 2007;35:142–145. - PubMed
- Lucas WJ. Plant viral movement proteins: Agents for cell-to-cell trafficking of viral genomes. Virology. 2006;344:169–184. - PubMed
- Radtke K, Dohner K, Sodeik B. Viral interactions with the cytoskeleton: A hitchhiker's guide to the cell. Cell Microbiol. 2006;8:387–400. - PubMed
- Greber UF, Way M. A superhighway to virus infection. Cell. 2006;124:741–754. - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources