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Sumoylation regulates the stability and nuclease activity of Saccharomyces cerevisiae Dna2

Published on Dec 1, 2019
路 DOI :10.1038/s42003-019-0428-0
Lepakshi Ranjha7
Estimated H-index: 7
(USI: University of Lugano),
Maryna Levikova7
Estimated H-index: 7
(UZH: University of Zurich)
+ 2 AuthorsPetr Cejka28
Estimated H-index: 28
(ETH Zurich)
Abstract
Dna2 is an essential nuclease-helicase that acts in several distinct DNA metabolic pathways including DNA replication and recombination. To balance these functions and prevent unscheduled DNA degradation, Dna2 activities must be regulated. Here we show that Saccharomyces cerevisiae Dna2 function is controlled by sumoylation. We map the sumoylation sites to the N-terminal regulatory domain of Dna2 and show that in vitro sumoylation of recombinant Dna2 impairs its nuclease but not helicase activity. In cells, the total levels of the non-sumoylatable Dna2 variant are elevated. However, non-sumoylatable Dna2 shows impaired nuclear localization and reduced recruitment to foci upon DNA damage. Non-sumoylatable Dna2 reduces the rate of DNA end resection, as well as impedes cell growth and cell cycle progression through S phase. Taken together, these findings show that in addition to Dna2 phosphorylation described previously, Dna2 sumoylation is required for the homeostasis of the Dna2 protein function to promote genome stability. Ranhja et al. find that budding yeast Dna2 is modified by sumoylation, and that sumoylation reduces the nuclease, but not helicase activity of Dna2 in vitro. In cells, expression of a Dna2 version that cannot be sumoylated leads to impaired DNA end resection and cell division.
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