PXD009734-1

PXD009734 is an original dataset announced via ProteomeXchange.

Title	Phosphoproteomic Analysis of Mec1/ATR Signaling in Response to Different Manners of Kinase Activation
Description	The Mec1/ATR kinase coordinates multiple cellular responses to replication stress. In addition to its canonical role in activating the checkpoint kinase Rad53, Mec1 also plays checkpoint-independent roles in genome maintenance that are not well understood. Here we employed a combined genetic-phosphoproteomic approach to manipulate Mec1 activation and globally monitor Mec1 signaling, allowing us to delineate distinct checkpoint-independent modes of Mec1 action. Using cells in which endogenous Mec1 activators were genetically ablated, we found that expression of “free” Mec1 Activation Domains (MADs) can robustly activate Mec1 and rescue the severe DNA replication and growth defects of these cells back to wild-type levels. However, unlike the activation mediated by endogenous activator-proteins, “free” MADs are unable to stimulate Mec1-mediated suppression of gross chromosomal rearrangements (GCRs), revealing that Mec1’s role in genome maintenance is separable from a previously unappreciated pro-replicative function. Both Mec1’s functions in promoting replication and suppressing GCRs are independent of the downstream checkpoint kinases. Additionally, Mec1-dependent GCR suppression seems to require localized Mec1 action at DNA lesions, which correlates with the phosphorylation of activator-proximal substrates involved in homologous recombination-mediated DNA repair. These findings establish that Mec1 initiates checkpoint signaling, promotes DNA replication, and maintains genetic stability through distinct modes of action.
HostingRepository	PRIDE
AnnounceDate	2019-01-14
AnnouncementXML	Submission_2019-01-14_02:57:48.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Michael Lanz
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	phosphorylated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2018-05-10 01:34:01	ID requested
⏵ 1	2019-01-14 02:57:49	announced

Lanz MC, Oberly S, Sanford EJ, Sharma S, Chabes A, Smolka MB, Separable roles for Mec1/ATR in genome maintenance, DNA replication, and checkpoint signaling. Genes Dev, 32(11-12):822-835(2018) [pubmed]

curator keyword: Biological

submitter keyword: Mec1, ATR, Phosphoproteomics, Dna2, Ddc1, Dpb11

Marcus B Smolka
contact affiliation	Department of Molecular Biology and Genetics, Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY 14853
contact email	mbs266@cornell.edu
lab head
Michael Lanz
contact affiliation	Cornell University
contact email	mcl246@cornell.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD009734
     1. Label: PRIDE project
     2. Name: Phosphoproteomic Analysis of Mec1/ATR Signaling in Response to Different Manners of Kinase Activation