Phospho-epitope binding by the BRCT domains of hPTIP controls multiple aspects of the cellular response to DNA damage.
Human (h)PTIP plays important but poorly understood roles in cellular responses to DNA damage. hPTIP interacts with 53BP1 tumour suppressor but only when 53BP1 is phosphorylated by ATM after DNA damage although the mechanism(s) and significance of the interaction of these two proteins are unclear. Here, we pinpoint a single ATM-phosphorylated residue in 53BP1--Ser25--that is required for binding of 53BP1 to hPTIP. Binding of phospho-Ser25 to hPTIP in vitro and in vivo requires two closely apposed pairs of BRCT domains at the C-terminus of hPTIP and neither pair alone can bind to phospho-Ser25, even though one of these BRCT pairs in isolation can bind to other ATM-phosphorylated epitopes. Mutations in 53BP1 and in hPTIP that prevent the interaction of the two proteins, render cells hypersensitive to DNA damage and weaken ATM signalling. The C-terminal BRCT domains of hPTIP are also required for stable retention of hPTIP at sites of DNA damage but this appears to be independent of binding to 53BP1. Thus, the BRCT domains of hPTIP play important roles in the cellular response to DNA damage.
Munoz
Ivan M
IM
MRC Protein Phosphorylation Unit, Sir James Black Centre, University of Dundee, Dundee DD1 5EH, Scotland, UK.
Jowsey
Paul A
PA
Toth
Rachel
R
Rouse
John
J
D064007:Ataxia Telangiectasia Mutated Proteins; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012694:Serine / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D000071857:Tumor Suppressor p53-Binding Protein 1
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; C485876:PAXIP1 protein, human; C498301:TP53BP1 protein, human; D025521:Tumor Suppressor Proteins; D000071857:Tumor Suppressor p53-Binding Protein 1; D010768:Phosphoserine; D012694:Serine; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases
Nucleic Acids Res. 2000 Jul 15;28(14):2741-51
10908331
Mol Cell. 2004 Dec 3;16(5):715-24
15574327
Curr Opin Cell Biol. 2001 Apr;13(2):225-31
11248557
J Biol Chem. 2001 Nov 9;276(45):42462-7
11571274
J Biol Chem. 2001 Dec 21;276(51):47759-62
11673449
Genes Dev. 2002 Jan 15;16(2):198-208
11799063
Science. 2002 Nov 15;298(5597):1435-8
12364621
Nat Cell Biol. 2002 Dec;4(12):998-1002
12447382
Mol Cell Biol. 2003 Mar;23(5):1666-73
12588986
Nat Rev Cancer. 2003 Mar;3(3):155-68
12612651
Mol Cell Biol. 2003 Apr;23(7):2556-63
12640136
J Biol Chem. 2003 Apr 25;278(17):14971-7
12578828
J Biol Chem. 2003 May 30;278(22):19579-82
12697768
J Biol Chem. 2003 Sep 19;278(38):36487-95
12824158
Science. 2003 Oct 24;302(5645):636-9
14576432
Science. 2003 Oct 24;302(5645):639-42
14576433
Cancer Res. 2004 Apr 1;64(7):2390-6
15059890
Nat Struct Mol Biol. 2004 Jun;11(6):512-8
15133502
Structure. 2004 Jul;12(7):1137-46
15242590
FASEB J. 1997 Jan;11(1):68-76
9034168
Cancer Res. 2004 Dec 15;64(24):9152-9
15604286
J Biol Chem. 2004 Dec 31;279(53):55562-9
15456759
Bioessays. 2005 Apr;27(4):397-407
15770685
Nature. 2005 Apr 14;434(7035):907-13
15829965
Radiother Oncol. 2005 Aug;76(2):119-22
16024119
Radiother Oncol. 2005 Aug;76(2):112-8
16026874
Mol Cell Biol. 2005 Nov;25(22):10079-86
16260621
J Med Chem. 2005 Dec 1;48(24):7829-46
16302822
Cell. 2005 Dec 29;123(7):1213-26
16377563
DNA Repair (Amst). 2006 Jun 10;5(6):741-9
16644291
J Cell Biol. 2000 Dec 25;151(7):1381-90
11134068