A 5′P degradation hot spot influences molecular farming of anticancerogenic nuclease TBN1 in tobacco cells

0464886 - BC 2017 RIV NL eng J - Článek v odborném periodiku
Týcová, Anna - Piernikarczyk, R.J.J. - Kugler, M. - Lipovová, P. - Podzimek, T. - Steger, G. - Matoušek, Jaroslav
A 5′P degradation hot spot influences molecular farming of anticancerogenic nuclease TBN1 in tobacco cells.
Plant Cell, Tissue and Organ Culture. Roč. 127, č. 2 (2016), s. 347-358. ISSN 0167-6857. E-ISSN 1573-5044
Institucionální podpora: RVO:60077344
Klíčová slova: mRNA quantification * Nicotiana benthamiana * Nicotiana tabacum * Post-transcriptional gene silencing
Kód oboru RIV: EB - Genetika a molekulární biologie
Impakt faktor: 2.002, rok: 2016

Tomato bifunctional nuclease 1 (TBN1) is a polyfunctional protein with anticancerogenic activity originally isolated as an overexpressed protein from viroid-infected tomato. Its molecular farming in plant cells could be a non-expensive source for its biotechnology preparation. So we analysed TBN1 expression in Agrobacterium-infiltrated leaf sectors of Nicotiana benthamiana and in transformed suspension culture of tobacco BY-2 cells. During its transient expression, TBN1 mRNA was strongly degraded within a hot spot localized in the 3′ region. This early degradation process was inhibited by PTGS suppressors p19 and p38 resulting in increased TBN1 mRNA and protein yield. In parallel to degradation of TBN1 mRNA, high mRNA levels of two RNA-dependent RNA polymerases were detected in infiltrated leaf sectors, as well as in the transformed tobacco suspension culture BY-2, where low expression of the nuclease was stably maintained. Higher TBN1 mRNA and nuclease activity levels were found during its molecular farming in RDR6-deficient N. benthamiana plants. By fluorescent microscopy of infiltrated and transformed plant cells, the nuclease-GFP fusion protein was shown to be organized in filament-like structures.
Trvalý link: http://hdl.handle.net/11104/0263635