S phase block following MEC1ATR inactivation occurs without severe dNTP depletion
2015 (English)In: Biology open, ISSN 2046-6390, Vol. 4, no 12, 1739-1743 p.Article in journal (Refereed) Published
Inactivation of Mec1, the budding yeast ATR, results in a permanent S phase arrest followed by chromosome breakage and cell death during G2/M. The S phase arrest is proposed to stem from a defect in Mec1-mediated degradation of Sml1, a conserved inhibitor of ribonucleotide reductase (RNR), causing a severe depletion in cellular dNTP pools. Here, the casual link between the S phase arrest, Sml1, and dNTP-levels is examined using a temperature sensitive mec1 mutant. In addition to S phase arrest, thermal inactivation of Mec1 leads to constitutively high levels of Sml1 and an S phase arrest. Expression of a novel suppressor, GIS2, a conserved mRNA binding zinc finger protein, rescues the arrest without down-regulating Sml1 levels. The dNTP pool in mec1 is reduced by ∼17% and GIS2 expression restores it, but only partially, to ∼93% of a control. We infer that the permanent S phase block following Mec1 inactivation can be uncoupled from its role in Sml1 down-regulation. Furthermore, unexpectedly modest effects of mec1 and GIS2 on dNTP levels suggest that the S phase arrest is unlikely to result from a severe depletion of dNTP pool as assumed, but a heightened sensitivity to small changes in its availability.
Place, publisher, year, edition, pages
2015. Vol. 4, no 12, 1739-1743 p.
Mec1, ATR, Sml1, RNR, dNTP, DNA replication, Replication arrest
Cell and Molecular Biology
IdentifiersURN: urn:nbn:se:umu:diva-111962DOI: 10.1242/bio.015347ISI: 000366672900016PubMedID: 26603472OAI: oai:DiVA.org:umu-111962DiVA: diva2:874451