Thioredoxin (Trxo1) Interacts with Proliferating Cell Nuclear Antigen (PCNA) and its Overexpression Affects the Growth of Tobacco Cell Culture

Calderón Pérez, Aingeru and Ortiz Espín, Ana María and Iglesias Fernandez, Raquel and Carbonero Zalduegui, Pilar and Pallardó, Federico Vicente and Sevilla, Francisca and Jimenez, Ana (2017). Thioredoxin (Trxo1) Interacts with Proliferating Cell Nuclear Antigen (PCNA) and its Overexpression Affects the Growth of Tobacco Cell Culture. "Redox Biology", v. 11 ; pp. 688-700. ISSN 2213-2317. https://doi.org/10.1016/j.redox.2017.01.018.

Description

Title: Thioredoxin (Trxo1) Interacts with Proliferating Cell Nuclear Antigen (PCNA) and its Overexpression Affects the Growth of Tobacco Cell Culture
Author/s:
  • Calderón Pérez, Aingeru
  • Ortiz Espín, Ana María
  • Iglesias Fernandez, Raquel
  • Carbonero Zalduegui, Pilar
  • Pallardó, Federico Vicente
  • Sevilla, Francisca
  • Jimenez, Ana
Item Type: Article
Título de Revista/Publicación: Redox Biology
Date: April 2017
Volume: 11
Subjects:
Faculty: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Biotecnología - Biología Vegetal
Creative Commons Licenses: Recognition - No derivative works - Non commercial

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Abstract

Thioredoxins (Trxs), key components of cellular redox regulation, act by controlling the redox status of many target proteins, and have been shown to play an essential role in cell survival and growth. The presence of a Trx system in the nucleus has received little attention in plants, and the nuclear targets of plant Trxs have not been conclusively identified. Thus, very little is known about the function of Trxs in this cellular compartment. Previously, we studied the intracellular localization of PsTrxo1 and confirmed its presence in mitochondria and, interestingly, in the nucleus under standard growth conditions. In investigating the nuclear function of PsTrxo1 we identified proliferating cellular nuclear antigen (PCNA) as a PsTrxo1 target by means of affinity chromatography techniques using purified nuclei from pea leaves. Such protein–protein interaction was corroborated by dot-blot and bimolecular fluorescence complementation (BiFC) assays, which showed that both proteins interact in the nucleus. Moreover, PsTrxo1 showed disulfide reductase activity on previously oxidized recombinant PCNA protein. In parallel, we studied the effects of PsTrxo1 overexpression on Tobacco Bright Yellow-2 (TBY-2) cell cultures. Microscopy and flow-cytometry analysis showed that PsTrxo1 overexpression increases the rate of cell proliferation in the transformed lines, with a higher percentage of the S phase of the cell cycle at the beginning of the cell culture (days 1 and 3) and at the G2/M phase after longer times of culture (day 9), coinciding with an upregulation of PCNA protein. Furthermore, in PsTrxo1 overexpressed cells there is a decrease in the total cellular glutathione content but maintained nuclear GSH accumulation, especially at the end of the culture, which is accompanied by a higher mitotic index, unlike non-overexpressing cells. These results suggest that Trxo1 is involved in the cell cycle progression of TBY-2 cultures, possibly through its link with cellular PCNA and glutathione.

Funding Projects

TypeCodeAcronymLeaderTitle
Government of SpainBFU2014-52452-PUnspecifiedUnspecifiedUnspecified
Government of SpainPROMETEOII/2014/056UnspecifiedUnspecifiedUnspecified
Government of SpainCSD2007-00057UnspecifiedUnspecifiedUnspecified

More information

Item ID: 53653
DC Identifier: http://oa.upm.es/53653/
OAI Identifier: oai:oa.upm.es:53653
DOI: 10.1016/j.redox.2017.01.018
Official URL: https://www.sciencedirect.com/science/article/pii/S2213231716303627
Deposited by: Biblioteca ETSI Agrónomos
Deposited on: 17 Jan 2019 13:53
Last Modified: 29 Apr 2019 14:18
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