Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds

DNA glycosylases initiate the base excision repair (BER) pathway by catalyzing the removal of damaged or mismatched bases from DNA. The Arabidopsis DNA glycosylase methyl-CpG-binding domain protein 4 like (MBD4L) is a nuclear enzyme triggering BER in response to the genotoxic agents 5-fluorouracil a...

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Main Authors: Lescano Lopez, Carlos Ignacio, Torres, José Roberto, Cecchini, Nicolás Miguel, Alvarez, María Elena
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: Wiley 2024
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/18506
https://onlinelibrary.wiley.com/doi/10.1111/tpj.16907
https://doi.org/10.1111/tpj.16907
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author Lescano Lopez, Carlos Ignacio
Torres, José Roberto
Cecchini, Nicolás Miguel
Alvarez, María Elena
author_browse Alvarez, María Elena
Cecchini, Nicolás Miguel
Lescano Lopez, Carlos Ignacio
Torres, José Roberto
author_facet Lescano Lopez, Carlos Ignacio
Torres, José Roberto
Cecchini, Nicolás Miguel
Alvarez, María Elena
author_sort Lescano Lopez, Carlos Ignacio
collection INTA Digital
description DNA glycosylases initiate the base excision repair (BER) pathway by catalyzing the removal of damaged or mismatched bases from DNA. The Arabidopsis DNA glycosylase methyl-CpG-binding domain protein 4 like (MBD4L) is a nuclear enzyme triggering BER in response to the genotoxic agents 5-fluorouracil and 5-bromouracil. To date, the involvement of MBD4L in plant physiological processes has not been analyzed. To address this, we studied the enzyme functions in seeds. We found that imbibition induced the MBD4L gene expression by generating two alternative transcripts, MBD4L.3 and MBD4L.4. Gene activation was stronger in aged than in non-aged seeds. Seeds from mbd4l-1 mutants displayed germination failures when maintained under control or ageing conditions, while 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 seeds reversed these phenotypes. Seed nuclear DNA repair, assessed by comet assays, was exacerbated in an MBD4L-dependent manner at 24 h post-imbibition. Under this condition, the BER genes ARP, APE1L, and LIG1 showed higher expression in 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 than in mbd4l-1 seeds, suggesting that these components could coordinate with MBD4L to repair damaged DNA bases in seeds. Interestingly, the ATM, ATR, BRCA1, RAD51, and WEE1 genes associated with the DNA damage response (DDR) pathway were activated in mbd4l-1, but not in 35S:MBD4L.3/mbd4l-1 or 35S:MBD4L.4/mbd4l-1 seeds. These results indicate that MBD4L is a key enzyme of a BER cascade that operates during seed imbibition, whose deficiency would cause genomic damage detected by DDR, generating a delay or reduction in germination.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
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spelling INTA185062024-07-15T12:20:58Z Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds Lescano Lopez, Carlos Ignacio Torres, José Roberto Cecchini, Nicolás Miguel Alvarez, María Elena Seed Germination DNA Damage Germinación de las Semillas Daño al ADN Base Excision Repair DNA Glycosylases Seed Ageing DNA Damage Responses MBD4L DNA glycosylases initiate the base excision repair (BER) pathway by catalyzing the removal of damaged or mismatched bases from DNA. The Arabidopsis DNA glycosylase methyl-CpG-binding domain protein 4 like (MBD4L) is a nuclear enzyme triggering BER in response to the genotoxic agents 5-fluorouracil and 5-bromouracil. To date, the involvement of MBD4L in plant physiological processes has not been analyzed. To address this, we studied the enzyme functions in seeds. We found that imbibition induced the MBD4L gene expression by generating two alternative transcripts, MBD4L.3 and MBD4L.4. Gene activation was stronger in aged than in non-aged seeds. Seeds from mbd4l-1 mutants displayed germination failures when maintained under control or ageing conditions, while 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 seeds reversed these phenotypes. Seed nuclear DNA repair, assessed by comet assays, was exacerbated in an MBD4L-dependent manner at 24 h post-imbibition. Under this condition, the BER genes ARP, APE1L, and LIG1 showed higher expression in 35S:MBD4L.3/mbd4l-1 and 35S:MBD4L.4/mbd4l-1 than in mbd4l-1 seeds, suggesting that these components could coordinate with MBD4L to repair damaged DNA bases in seeds. Interestingly, the ATM, ATR, BRCA1, RAD51, and WEE1 genes associated with the DNA damage response (DDR) pathway were activated in mbd4l-1, but not in 35S:MBD4L.3/mbd4l-1 or 35S:MBD4L.4/mbd4l-1 seeds. These results indicate that MBD4L is a key enzyme of a BER cascade that operates during seed imbibition, whose deficiency would cause genomic damage detected by DDR, generating a delay or reduction in germination. Instituto de Fisiología y Recursos Genéticos Vegetales Fil: Lescano Lopez, Carlos Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Estudios Agropecuarios (UDEA); Argentina Fil: Lescano Lopez, Carlos Ignacio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Fisiología y Recursos Genéticos Vegetales; Argentina Fil: Lescano Lopez, Carlos Ignacio. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas; Argentina Fil: Torres, José Roberto. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC). Departamento de Química Biológica Ranwel Caputto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cecchini, Nicolás Miguel. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC). Departamento de Química Biológica Ranwel Caputto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Alvarez, María Elena. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC). Departamento de Química Biológica Ranwel Caputto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2024-07-15T11:40:58Z 2024-07-15T11:40:58Z 2024-07-04 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/18506 https://onlinelibrary.wiley.com/doi/10.1111/tpj.16907 0960-7412 1365-313X (online) https://doi.org/10.1111/tpj.16907 eng info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Wiley The Plant Journal : 1-12 (First published: 04 July 2024)
spellingShingle Seed Germination
DNA Damage
Germinación de las Semillas
Daño al ADN
Base Excision Repair
DNA Glycosylases
Seed Ageing
DNA Damage Responses
MBD4L
Lescano Lopez, Carlos Ignacio
Torres, José Roberto
Cecchini, Nicolás Miguel
Alvarez, María Elena
Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds
title Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds
title_full Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds
title_fullStr Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds
title_full_unstemmed Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds
title_short Arabidopsis DNA glycosylase MBD4L improves recovery of aged seeds
title_sort arabidopsis dna glycosylase mbd4l improves recovery of aged seeds
topic Seed Germination
DNA Damage
Germinación de las Semillas
Daño al ADN
Base Excision Repair
DNA Glycosylases
Seed Ageing
DNA Damage Responses
MBD4L
url http://hdl.handle.net/20.500.12123/18506
https://onlinelibrary.wiley.com/doi/10.1111/tpj.16907
https://doi.org/10.1111/tpj.16907
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