RNAi technology

Komarova, T.V.; Citovsky, V.; Dorokhov, Y.L.

Авторы: Komarova T.V., Citovsky V., Dorokhov Y.L.
Год издания: 2011
Место издания: CRC Press, Taylor CRC Press, Taylor & Francis Group, USA Science, Technology, and Medical eBooks from Taylor & Francis Group Taylor & Francis Group 6000 Broken Sound Parkway,NW Suite 300 Boca Raton, FL 33487
Объём: 364 страниц
ISBN: 978-1-4398-7325-0
DOI: 10.1201/b10882-8
Аннотация: A ubiquitous cell wall enzyme, pectin methylesterase (PME), a multifunctional protein which catalyzes pectin deesterification, producing atmospheric methanol, participates in cell wall modulation during general plant growth and pollen tube growth. The tobacco PME interacts with the movement protein of Tobacco mosaic virus (TMV) suggesting that PME may be involved in cell-to-cell movement of plant viruses, whereas methanol produced by PME-catalyzed reactions may act as a signal molecule in plant-plant and plant-pathogen interactions. Furthermore, PME also is an efficient enhancer of virus- and transgene-induced gene silencing (VIGS and TIGS, respectively) via activation of siRNA and miRNA production. Enhancement of VIGS and TIGS is accompanied by relocation of the DCL1 protein from the cell nucleus to the cytoplasm. This chapter shows that PME can prevent nuclear transport of reporter proteins, GFP and GFP-tagged Tobacco mosaic virus movement protein and MS2 phage coat protein fused with a nuclear localization signal (NLS). Furthermore, PME reduced nuclear transport of a plant protein ALY. Interestingly, this PME activity enhanced the effect of a plant virus RNA silencing suppressor, the Tomato bushy stunt virus P19 protein, which is known to relocate the host ALY to the cell cytoplasm. PME production is elevated under stress conditions, and the invading virus may exploit the ability of PME to impair ALY nuclear transport for augmenting the activity of its P19 suppressor. Indeed, coexpression of PME and P19 substantially enhanced TMV infection. The crucifer-infecting TMV (crTMV) RNA accumulation was increased in 10 times at 3 dpi in leaf areas co-agroinjected with P19 and PME. The GFP production in leaves infiltrated with crTMV:GFP together with binary vectors expressing PME and P19 was doubled in comparison with P19 alone. Because only enzymatically active PME, which is secreted into cell wall, was able to suppress nuclear import, we hypothesize that the products of pectin deesterification, demethylesterified homogalacturonan or methanol, may immediate the effects of PME on nuclear import.
Добавил в систему: Дорохов Юрий Леонидович

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