Laser microirradiation as a versatile system for probing protein recruitment and protein-protein interactions at DNA lesions in plants
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F22%3A00127898" target="_blank" >RIV/00216224:14740/22:00127898 - isvavai.cz</a>
Result on the web
<a href="https://nph.onlinelibrary.wiley.com/doi/10.1111/nph.18086" target="_blank" >https://nph.onlinelibrary.wiley.com/doi/10.1111/nph.18086</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1111/nph.18086" target="_blank" >10.1111/nph.18086</a>
Alternative languages
Result language
angličtina
Original language name
Laser microirradiation as a versatile system for probing protein recruitment and protein-protein interactions at DNA lesions in plants
Original language description
Plant protoplasts are generated by treatment with digestion enzymes, producing plant cells devoid of the cell wall and competent for efficient polyethylene glycol mediated transformation. This way fluorescently tagged proteins can be introduced to the protoplasts creating an excellent system to probe the localization and function of uncharacterized plant proteins in vivo. We implement the method of laser microirradiation to generate DNA lesions in Arabidopsis thaliana, which enables monitoring the recruitment and dynamics of the DNA repair factors as well as bimolecular fluorescence complementation assay to test transient, conditional interactions of proteins directly at sites of DNA damage. We demonstrate that laser microirradiation in protoplasts yields a physiological cellular response to DNA lesions, based on proliferating cell nuclear antigen (PCNA) redistribution in the nucleus and show that factors involved in DNA repair, such as MRE11 or PCNA are recruited to induced DNA lesions. This technique is relatively easy to adopt by other laboratories and extends the current toolkit of methods aimed to understand the details of DNA damage response in plants. The presented method is fast, flexible and facilitates work with different mutant backgrounds or even different species, extending the utility of the system.
Czech name
—
Czech description
—
Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
—
OECD FORD branch
10611 - Plant sciences, botany
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2022
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
New Phytologist
ISSN
0028-646X
e-ISSN
—
Volume of the periodical
234
Issue of the periodical within the volume
5
Country of publishing house
US - UNITED STATES
Number of pages
10
Pages from-to
1891-1900
UT code for WoS article
000785919300001
EID of the result in the Scopus database
2-s2.0-85129048753