PMID- 26929199
OWN - NLM
STAT- MEDLINE
DCOM- 20160906
LR  - 20181113
IS  - 1098-5549 (Electronic)
IS  - 0270-7306 (Print)
IS  - 0270-7306 (Linking)
VI  - 36
IP  - 9
DP  - 2016 May
TI  - Phosphorylation-Dependent Regulation of the DNA Damage Response of Adaptor 
      Protein KIBRA in Cancer Cells.
PG  - 1354-65
LID - 10.1128/MCB.01004-15 [doi]
AB  - Multifunctional adaptor proteins encompassing various protein-protein interaction 
      domains play a central role in the DNA damage response pathway. In this report, 
      we show that KIBRA is a physiologically interacting reversible substrate of 
      ataxia telangiectasia mutated (ATM) kinase. We identified the site of 
      phosphorylation in KIBRA as threonine 1006, which is embedded within the 
      serine/threonine (S/T) Q consensus motif, by site-directed mutagenesis, and we 
      further confirmed the same with a phospho-(S/T) Q motif-specific antibody. 
      Results from DNA repair functional assays such as the gamma-H2AX assay, pulsed-field 
      gel electrophoresis (PFGE), Comet assay, terminal 
      deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay, 
      and clonogenic cell survival assay using stable overexpression clones of 
      wild-type (wt.) KIBRA and active (T1006E) and inactive (T1006A) KIBRA 
      phosphorylation mutants showed that T1006 phosphorylation on KIBRA is essential 
      for optimal DNA double-strand break repair in cancer cells. Further, results from 
      stable retroviral short hairpin RNA-mediated knockdown (KD) clones of KIBRA and 
      KIBRA knockout (KO) model cells generated by a clustered regularly interspaced 
      short palindromic repeat (CRISPR)-Cas9 system showed that depleting KIBRA levels 
      compromised the DNA repair functions in cancer cells upon inducing DNA damage. 
      All these phenotypic events were reversed upon reconstitution of KIBRA into cells 
      lacking KIBRA knock-in (KI) model cells. All these results point to the fact that 
      phosphorylated KIBRA might be functioning as a scaffolding protein/adaptor 
      protein facilitating the platform for further recruitment of other DNA damage 
      response factors. In summary, these data demonstrate the imperative functional 
      role of KIBRAper se(KIBRA phosphorylation at T1006 site as a molecular switch 
      that regulates the DNA damage response, possibly via the nonhomologous end 
      joining [NHEJ] pathway), suggesting that KIBRA could be a potential therapeutic 
      target for modulating chemoresistance in cancer cells.
CI  - Copyright (c) 2016, American Society for Microbiology. All Rights Reserved.
FAU - Mavuluri, Jayadev
AU  - Mavuluri J
AD  - Department of Biotechnology, Indian Institute of Technology Madras (IITM), 
      Chennai, India.
FAU - Beesetti, Swarnalatha
AU  - Beesetti S
AD  - Department of Biotechnology, Indian Institute of Technology Madras (IITM), 
      Chennai, India.
FAU - Surabhi, Rohan
AU  - Surabhi R
AD  - Department of Biotechnology, Indian Institute of Technology Madras (IITM), 
      Chennai, India Departments of Human Genetics, Sri Ramachandra University, Porur, 
      Chennai, India.
FAU - Kremerskothen, Joachim
AU  - Kremerskothen J
AD  - Department of Nephrology, Hypertension and Rheumatology, University Hospital 
      Muenster, Muenster, Germany.
FAU - Venkatraman, Ganesh
AU  - Venkatraman G
AD  - Departments of Human Genetics, Sri Ramachandra University, Porur, Chennai, India 
      ganeshv@sriramachandra.edu.in rayala@iitm.ac.in.
FAU - Rayala, Suresh K
AU  - Rayala SK
AD  - Department of Biotechnology, Indian Institute of Technology Madras (IITM), 
      Chennai, India ganeshv@sriramachandra.edu.in rayala@iitm.ac.in.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160415
PL  - United States
TA  - Mol Cell Biol
JT  - Molecular and cellular biology
JID - 8109087
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (WWC1 protein, human)
RN  - 11056-06-7 (Bleomycin)
RN  - EC 2.7.11.1 (ATM protein, human)
RN  - EC 2.7.11.1 (Ataxia Telangiectasia Mutated Proteins)
SB  - IM
MH  - Ataxia Telangiectasia Mutated Proteins/genetics/metabolism
MH  - Bleomycin/pharmacology
MH  - Breast Neoplasms/*genetics/metabolism/pathology
MH  - Cell Line, Tumor
MH  - DNA Breaks, Double-Stranded
MH  - DNA Damage/physiology
MH  - DNA Repair/*physiology
MH  - Drug Resistance, Neoplasm/drug effects/genetics
MH  - Female
MH  - Gene Knockout Techniques
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins/genetics/*metabolism
MH  - Phosphoproteins/genetics/*metabolism
MH  - Phosphorylation
PMC - PMC4836221
EDAT- 2016/03/02 06:00
MHDA- 2016/09/07 06:00
PMCR- 2016/10/15
CRDT- 2016/03/02 06:00
PHST- 2015/11/09 00:00 [received]
PHST- 2016/02/12 00:00 [accepted]
PHST- 2016/03/02 06:00 [entrez]
PHST- 2016/03/02 06:00 [pubmed]
PHST- 2016/09/07 06:00 [medline]
PHST- 2016/10/15 00:00 [pmc-release]
AID - MCB.01004-15 [pii]
AID - 01004-15 [pii]
AID - 10.1128/MCB.01004-15 [doi]
PST - epublish
SO  - Mol Cell Biol. 2016 Apr 15;36(9):1354-65. doi: 10.1128/MCB.01004-15. Print 2016 
      May.