The topoisomerase 3α zinc-finger domain T1 of Arabidopsis thaliana is required for targeting the enzyme activity to Holliday junction-like DNA repair intermediates

PLoS Genetics
Annika DornHolger Puchta

Abstract

Topoisomerase 3α, a class I topoisomerase, consists of a TOPRIM domain, an active centre and a variable number of zinc-finger domains (ZFDs) at the C-terminus, in multicellular organisms. Whereas the functions of the TOPRIM domain and the active centre are known, the specific role of the ZFDs is still obscure. In contrast to mammals where a knockout of TOP3α leads to lethality, we found that CRISPR/Cas induced mutants in Arabidopsis are viable but show growth retardation and meiotic defects, which can be reversed by the expression of the complete protein. However, complementation with AtTOP3α missing either the TOPRIM-domain or carrying a mutation of the catalytic tyrosine of the active centre leads to embryo lethality. Surprisingly, this phenotype can be overcome by the simultaneous removal of the ZFDs from the protein. In combination with a mutation of the nuclease AtMUS81, the TOP3α knockout proved to be also embryo lethal. Here, expression of TOP3α without ZFDs, and in particular without the conserved ZFD T1, leads to only a partly complementation in root growth-in contrast to the complete protein, that restores root length to mus81-1 mutant level. Expressing the E. coli resolvase RusA in this background, which is able to p...Continue Reading

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Citations

Aug 6, 2019·The Plant Journal : for Cell and Molecular Biology·Felix Wolter, Holger Puchta
Apr 12, 2020·The Journal of Biological Chemistry·Anna H Bizard, Ian D Hickson
Dec 15, 2019·Genes·Annika Dorn, Holger Puchta
Sep 6, 2019·International Journal of Molecular Sciences·Janina EnderleHolger Puchta
Oct 22, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Tumpa DasguptaYuk-Ching Tse-Dinh
Sep 16, 2020·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Marta GòdiaAlex Clop
Dec 29, 2020·Structure·Rohan Bythell-Douglas, Andrew J Deans
Feb 24, 2021·The Plant Journal : for Cell and Molecular Biology·Amy Leanne WhitbreadHolger Puchta
May 8, 2021·Nature Plants·Michelle RönspiesHolger Puchta

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Datasets Mentioned

BETA
GSE116582

Methods Mentioned

BETA
RNA-seq
transgenic
PCR
light microscopy
genotyping

Software Mentioned

SmartRoot
CLC Genomics Workbench
ImageJ

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