Tolerance to replication stress requires Dun1p kinase and activation of the electron transport chain

Nagar, Shreya; Mehta, Riddhi; Kaur, Pritpal; Liliah, Roshini T.; Vančura, Aleš

doi:10.1016/j.bbamcr.2022.119382

Cited by 1 publication

(3 citation statements)

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“…Cell cycle arrest in G1 phase by α-factor and release into S phase was carried out as described 25 , 37 , 73 , 74 by adding α-factor to 10 µg/ml to cells exponentially growing in YPD medium. Following α-factor addition, the cultures were incubated for 2 h, and an additional 5 µg/ml of α-factor was added and incubation continued for another 1 h. The culture was divided in 2 aliquots, and one aliquot treated for 30 min with HU for the last 30 min of the α-factor arrest.…”

Section: Methodsmentioning

confidence: 99%

“…The HU treatment of S-phase cells was performed by arresting WT (W303-1a) cells with α-factor, washing them 2 times with YPD medium pre-warmed to 30 °C, and releasing them into YPD medium or YPD medium containing HU for 30 min. The cell cycle arrest in G1 phase and release into S phase was monitored by flow cytometry of Sytox Green stained cells using Sigma Millipore Guava easy Cyte flow cytometer as described 74 . This protocol yields about 90% of cells arrested in G1 phase and released into S phase 74 .…”

Section: Methodsmentioning

confidence: 99%

“…The cell cycle arrest in G1 phase and release into S phase was monitored by flow cytometry of Sytox Green stained cells using Sigma Millipore Guava easy Cyte flow cytometer as described 74 . This protocol yields about 90% of cells arrested in G1 phase and released into S phase 74 .…”

Section: Methodsmentioning

confidence: 99%

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Hydroxyurea and inactivation of checkpoint kinase MEC1 inhibit transcription termination and pre-mRNA cleavage at polyadenylation sites in budding yeast

Kaur,

Nagar,

Mehta

et al. 2023

Sci Rep

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The DNA damage response (DDR) is an evolutionarily conserved process essential for cell survival. The transcription changes triggered by DDR depend on the nature of DNA damage, activation of checkpoint kinases, and the stage of cell cycle. The transcription changes can be localized and affect only damaged DNA, but they can be also global and affect genes that are not damaged. While the purpose of localized transcription inhibition is to avoid transcription of damaged genes and make DNA accessible for repair, the purpose and mechanisms of global transcription inhibition of undamaged genes are less well understood. We show here that a brief cell treatment with hydroxyurea (HU) globally inhibits RNA synthesis and transcription by RNA polymerase I, II, and III (RNAPI, RNAPII, and RNAPIII). HU reduces efficiency of transcription termination and inhibits pre-mRNA cleavage at the polyadenylation (pA) sites, destabilizes mRNAs, and shortens poly(A) tails of mRNAs, indicating defects in pre-mRNA 3′ end processing. Inactivation of the checkpoint kinase Mec1p downregulates the efficiency of transcription termination and reduces the efficiency of pre-mRNAs clevage at the pA sites, suggesting the involvement of DNA damage checkpoint in transcription termination and pre-mRNA 3′ end processing.

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