PXD024768

PXD024768 is an original dataset announced via ProteomeXchange.

Title	The Conserved Tpk1 Regulates Non-Homologous End Joining Double-Strand Break Repair by Phosphorylation of Nej1, a Homolog of the Human XLF
Description	The yeast cyclic-AMP dependent protein kinase A (PKA) is a ubiquitous serine-threonine kinase, encompassing three catalytic (Tpk1-3) and one regulatory (Bcy1) subunits. Evidence suggests PKA involvement in DNA damage checkpoint response, but how DNA repair pathways are regulated by PKA subunits remains inconclusive. Here, we report that deleting the catalytic subunit, tpk1, reduces non-homologous end joining (NHEJ) efficiency, whereas tpk2-3 and bcy1 deletion does not. Epistatic analyses revealed that tpk1, as well as the DNA damage checkpoint kinase (dun1) and NHEJ regulator (nej1), co-function in the same pathway. Chromatin immunoprecipitation and resection data suggest that the recruitment of repair proteins and DNA resection is influenced by tpk1 deletion. Further, we show that Tpk1 phosphorylation of Nej1 at S298 (a Dun1 phosphosite) is indispensable for NHEJ repair, in addition to nuclear targeting of Nej1 and its binding partner Lif1. In mammalian cells, loss of PRKACB (the human homolog of Tpk1) also reduced NHEJ efficiency, and similarly, PRKACB was found to phosphorylate XLF (a Nej1 human homolog) at S263, a corresponding residue of the yeast Nej1 S298. Together, our results uncover a new and conserved mechanism for Tpk1 and PRKACB in phosphorylating Nej1 (or XLF), which is critically required for NHEJ repair.
HostingRepository	PRIDE
AnnounceDate	2022-02-17
AnnouncementXML	Submission_2022-02-16_23:16:20.875.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sadhna Phanse
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	mono N-acetylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2021-03-16 04:58:10	ID requested
⏵ 1	2022-02-16 23:16:21	announced

Jessulat M, Amin S, Hooshyar M, Malty R, Moutaoufik MT, Zilocchi M, Istace Z, Phanse S, Aoki H, Omidi K, Burnside D, Samanfar B, Aly KA, Golshani A, Babu M, The conserved Tpk1 regulates non-homologous end joining double-strand break repair by phosphorylation of Nej1, a homolog of the human XLF. Nucleic Acids Res, 49(14):8145-8160(2021) [pubmed]

submitter keyword: DSB repair, Genomic instability, Kinase, NHEJ, NEJ1, PRKACB, TPK1

Mohan Babu
contact affiliation	Department of Biochemistry, University of Regina, Regina, Saskatchewan S4S 0A2, Canada.
contact email	mohan.babu@uregina.ca
lab head
Sadhna Phanse
contact affiliation	University of Toronto; University of Regina
contact email	srphanse@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD024768
     1. Label: PRIDE project
     2. Name: The Conserved Tpk1 Regulates Non-Homologous End Joining Double-Strand Break Repair by Phosphorylation of Nej1, a Homolog of the Human XLF