Ser612

Site Information
MyLsPVRsPKKKGsT SwissProt Entrez-Gene
Blast this site against: NCBI SwissProt PDB
Site Group ID: 447977

In vivo Characterization
Methods used to characterize site in vivo:	[32P] bio-synthetic labeling ( 52 ) , electrophoretic mobility shift ( 52 ) , flow cytometry ( 51 ) , immunoprecipitation ( 47 ) , mass spectrometry ( 1 , 2 , 4 , 5 , 6 , 8 , 9 , 11 , 12 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 48 ) , mass spectrometry (in vitro) ( 22 ) , mutation of modification site ( 7 , 10 , 47 , 52 ) , phospho-antibody ( 3 , 7 , 47 , 50 , 51 ) , western blotting ( 3 , 7 , 10 , 47 , 51 )
Disease tissue studied:	bone cancer ( 3 , 7 ) , brain cancer ( 35 ) , glioblastoma ( 35 ) , glioma ( 35 ) , breast cancer ( 2 , 6 , 11 , 51 ) , HER2 positive breast cancer ( 1 ) , luminal A breast cancer ( 1 ) , luminal B breast cancer ( 1 ) , breast cancer, triple negative ( 1 ) , cervical cancer ( 7 , 10 , 52 ) , leukemia ( 7 , 14 , 48 , 50 ) , acute lymphocytic leukemia ( 50 ) , acute myelogenous leukemia ( 14 ) , chronic myelogenous leukemia ( 7 , 48 ) , T cell leukemia ( 7 ) , lung cancer ( 11 , 36 ) , non-small cell lung cancer ( 11 , 36 ) , lymphoma ( 7 ) , neuroblastoma ( 7 ) , melanoma skin cancer ( 4 )
Relevant cell line - cell type - tissue:	293 (epithelial) [AT1 (human), transfection, AT1R stable transfected HEK293] ( 21 ) , A549 (pulmonary) ( 8 ) , bone marrow ( 50 ) , breast ( 1 ) , C33-A (cervical) ( 7 , 10 , 52 ) , Calu 6 (pulmonary) ( 11 ) , Flp-In T-Rex-293 (epithelial) [PRKD1 (human), genetic knockin] ( 12 ) , Flp-In T-Rex-293 (epithelial) ( 12 ) , H2009 (pulmonary) ( 11 ) , H2077 (pulmonary) ( 11 ) , H2887 (pulmonary) ( 11 ) , H322M (pulmonary) ( 11 ) , HCC1359 (pulmonary) ( 11 ) , HCC1937 (breast cell) ( 11 ) , HCC2279 (pulmonary) ( 11 ) , HCC4006 (pulmonary) ( 11 ) , HCT15 (intestinal) ( 47 ) , HeLa (cervical) ( 5 ) , HFFF-2 (fibroblast) ( 52 ) , HMLER ('stem, breast cancer') [CXCR4 (human), knockdown] ( 6 ) , HMLER ('stem, breast cancer') ( 6 ) , HOP62 (pulmonary) ( 11 ) , HUES-9 ('stem, embryonic') ( 20 ) , Jurkat (T lymphocyte) ( 9 , 15 , 16 , 17 , 18 , 19 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 ) , K562 (erythroid) ( 7 , 48 ) , KG-1 (myeloid) ( 14 ) , LOU-NH91 (squamous) ( 11 ) , lung ( 36 ) , MCF-7 (breast cell) ( 2 , 11 , 47 , 51 ) , MKN-45 (gastric) ( 37 , 38 ) , MOLT-4 (T lymphocyte) ( 7 , 47 ) , NCI-H1395 (pulmonary) ( 11 ) , NCI-H1648 (pulmonary) ( 11 ) , NCI-H2030 (pulmonary) ( 11 ) , NCI-H2172 (pulmonary) ( 11 ) , NCI-H322 (pulmonary) ( 11 ) , NCI-H460 (pulmonary) ( 11 ) , Rat1 (fibroblast) ( 52 ) , Saos-2 (bone cell) [Rb (human)] ( 47 ) , Saos-2 (bone cell) ( 3 , 7 ) , SH-SY5Y (neural crest) ( 7 ) , SKBr3 (breast cell) ( 51 ) , T47D (breast cell) ( 51 ) , U-937 (myeloid) ( 7 ) , U87MG (glial) ( 35 ) , WM239A (melanocyte) ( 4 )

Upstream Regulation
Regulatory protein:	ATM (human) ( 47 ) , Chk1 (human) ( 47 ) , Chk2 (human) ( 47 ) , EGFR (human) ( 35 )
Putative in vivo kinases:	Chk2 (human) ( 47 ) , UL97 (herpesvirus) ( 3 )
Kinases, in vitro:	CDK2 (human) ( 49 , 50 ) , CDK4 (human) ( 49 ) , CDK6 (human) ( 49 ) , Chk1 (human) ( 47 ) , Chk2 (human) ( 47 )
Treatments:	actinomycin_D ( 47 ) , caffeine ( 47 ) , Chk2_inhibitor_II ( 7 ) , differentiation ( 7 ) , etoposide ( 7 ) , GW_8510 ( 7 ) , HPR ( 51 ) , ionizing_radiation ( 47 ) , KU-55933 ( 47 ) , phorbol_ester ( 7 ) , SB218078 ( 47 ) , siRNA ( 47 ) , UV ( 47 )

Downstream Regulation
Effects of modification on Rb:	molecular association, regulation ( 7 , 22 , 47 , 52 ) , protein conformation ( 22 )
Effects of modification on biological processes:	apoptosis, induced ( 47 ) , cell differentiation, induced ( 7 ) , transcription, altered ( 47 )
Induce interaction with:	E2F1 (human) ( 7 , 47 )
Inhibit interaction with:	E2F1 (human) ( 22 , 52 )

References
1	Mertins P, et al. (2016) Proteogenomics connects somatic mutations to signalling in breast cancer. Nature 534, 55-62 27251275 Curated Info
2	Carrier M, et al. (2016) Phosphoproteome and Transcriptome of RA-Responsive and RA-Resistant Breast Cancer Cell Lines. PLoS One 11, e0157290 27362937 Curated Info
3	Iwahori S, Hakki M, Chou S, Kalejta RF (2015) Molecular Determinants for the Inactivation of the Retinoblastoma Tumor Suppressor by the Viral Cyclin-dependent Kinase UL97. J Biol Chem 290, 19666-80 26100623 Curated Info
4	Stuart SA, et al. (2015) A Phosphoproteomic Comparison of B-RAFV600E and MKK1/2 Inhibitors in Melanoma Cells. Mol Cell Proteomics 14, 1599-615 25850435 Curated Info
5	Sharma K, et al. (2014) Ultradeep human phosphoproteome reveals a distinct regulatory nature of Tyr and Ser/Thr-based signaling. Cell Rep 8, 1583-94 25159151 Curated Info
6	Yi T, et al. (2014) Quantitative phosphoproteomic analysis reveals system-wide signaling pathways downstream of SDF-1/CXCR4 in breast cancer stem cells. Proc Natl Acad Sci U S A 111, E2182-90 24782546 Curated Info
7	Hattori T, et al. (2014) Distinct and site-specific phosphorylation of the retinoblastoma protein at serine 612 in differentiated cells. PLoS One 9, e86709 24466208 Curated Info
8	Kim JY, et al. (2013) Dissection of TBK1 signaling via phosphoproteomics in lung cancer cells. Proc Natl Acad Sci U S A 110, 12414-9 23836654 Curated Info
9	Mertins P, et al. (2013) Integrated proteomic analysis of post-translational modifications by serial enrichment. Nat Methods 10, 634-7 23749302 Curated Info
10	Lentine B, et al. (2012) Dephosphorylation of threonine-821 of the retinoblastoma tumor suppressor protein (Rb) is required for apoptosis induced by UV and Cdk inhibition. Cell Cycle 11, 3324-30 22895174 Curated Info
11	Klammer M, et al. (2012) Phosphosignature predicts dasatinib response in non-small cell lung cancer. Mol Cell Proteomics 11, 651-68 22617229 Curated Info
12	Franz-Wachtel M, et al. (2012) Global detection of protein kinase D-dependent phosphorylation events in nocodazole-treated human cells. Mol Cell Proteomics 11, 160-70 22496350 Curated Info
13	Carnevale J, Palander O, Seifried LA, Dick FA (2012) DNA damage signals through differentially modified E2F1 molecules to induce apoptosis. Mol Cell Biol 32, 900-12 22184068 Curated Info
14	Weber C, Schreiber TB, Daub H (2012) Dual phosphoproteomics and chemical proteomics analysis of erlotinib and gefitinib interference in acute myeloid leukemia cells. J Proteomics 75, 1343-56 22115753 Curated Info
15	Mulhern D (2011) CST Curation Set: 12677; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (F/Y/M)Xp[ST](L/I/M) Curated Info
16	Guo A (2011) CST Curation Set: 12438; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
17	Guo A (2011) CST Curation Set: 12068; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
18	Guo A (2011) CST Curation Set: 11986; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]XP Curated Info
19	Guo A (2011) CST Curation Set: 11987; Year: 2011; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]XP Curated Info
20	Rigbolt KT, et al. (2011) System-wide temporal characterization of the proteome and phosphoproteome of human embryonic stem cell differentiation. Sci Signal 4, rs3 21406692 Curated Info
21	Christensen GL, et al. (2010) Quantitative phosphoproteomics dissection of seven-transmembrane receptor signaling using full and biased agonists. Mol Cell Proteomics 9, 1540-53 20363803 Curated Info
22	Burke JR, Deshong AJ, Pelton JG, Rubin SM (2010) Phosphorylation-induced conformational changes in the retinoblastoma protein inhibit E2F transactivation domain binding. J Biol Chem 285, 16286-93 20223825 Curated Info
23	Possemato A (2010) CST Curation Set: 9695; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: YXp[ST] Curated Info
24	Possemato A (2010) CST Curation Set: 9696; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: YXp[ST] Curated Info
25	Possemato A (2010) CST Curation Set: 9642; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pSPX(I/L/M/V) Curated Info
26	Possemato A (2010) CST Curation Set: 9284; Year: 2010; Biosample/Treatment: cell line, Jurkat/pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
27	Possemato A (2010) CST Curation Set: 9286; Year: 2010; Biosample/Treatment: cell line, Jurkat/pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Curated Info
28	Possemato A (2010) CST Curation Set: 9288; Year: 2010; Biosample/Treatment: cell line, Jurkat/pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
29	Possemato A (2010) CST Curation Set: 9066; Year: 2010; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (F/Y)XpS Curated Info
30	Zhou J (2009) CST Curation Set: 6956; Year: 2009; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: YXp[ST] Curated Info
31	Zhou J (2009) CST Curation Set: 6052; Year: 2009; Biosample/Treatment: cell line, Jurkat/untreated; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (F/Y/M)Xp[ST](L/I/M) Curated Info
32	Zhou J (2009) CST Curation Set: 6054; Year: 2009; Biosample/Treatment: cell line, Jurkat/TPA; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (F/Y/M)Xp[ST](L/I/M) Curated Info
33	Zhou J (2009) CST Curation Set: 6053; Year: 2009; Biosample/Treatment: cell line, Jurkat/TPA; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (F/Y/M)Xp[ST](L/I/M) Curated Info
34	Zhou J (2009) CST Curation Set: 6051; Year: 2009; Biosample/Treatment: cell line, Jurkat/untreated; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: (F/Y/M)Xp[ST](L/I/M) Curated Info
35	Joughin BA, et al. (2009) An integrated comparative phosphoproteomic and bioinformatic approach reveals a novel class of MPM-2 motifs upregulated in EGFRvIII-expressing glioblastoma cells. Mol Biosyst 5, 59-67 19081932 Curated Info
36	Rikova K (2008) CST Curation Set: 5536; Year: 2008; Biosample/Treatment: tissue, lung/untreated; Disease: non-small cell lung cancer; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: pY Antibodies Used to Purify Peptides prior to LCMS: Phospho-Tyrosine Mouse mAb (P-Tyr-100) Cat#: 9411, PTMScan(R) Phospho-Tyr Motif (Y*) Immunoaffinity Beads Cat#: 1991 Curated Info
37	Guo A (2008) CST Curation Set: 5570; Year: 2008; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
38	Guo A (2008) CST Curation Set: 5571; Year: 2008; Biosample/Treatment: cell line, MKN-45/Su11274; Disease: gastric carcinoma; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
39	Zhou J (2008) CST Curation Set: 4704; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
40	Possemato A (2008) CST Curation Set: 4344; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
41	Possemato A (2008) CST Curation Set: 4345; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, Phospho-MAPK/CDK Substrates (PXSP or SPXR/K) (34B2) Rabbit mAb Cat#: 2325, Phospho-(Ser/Thr) PKD Substrate Antibody Cat#: 4381 Curated Info
42	Possemato A (2008) CST Curation Set: 4346; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, Phospho-MAPK/CDK Substrates (PXSP or SPXR/K) (34B2) Rabbit mAb Cat#: 2325, Phospho-(Ser/Thr) PKD Substrate Antibody Cat#: 4381 Curated Info
43	Possemato A (2008) CST Curation Set: 4347; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, Phospho-MAPK/CDK Substrates (PXSP or SPXR/K) (34B2) Rabbit mAb Cat#: 2325, Phospho-(Ser/Thr) PKD Substrate Antibody Cat#: 4381 Curated Info
44	Zhou J (2008) CST Curation Set: 3678; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
45	Zhou J (2008) CST Curation Set: 3679; Year: 2008; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
46	Possemato A (2007) CST Curation Set: 3480; Year: 2007; Biosample/Treatment: cell line, Jurkat/calyculin_A & pervanadate; Disease: T cell leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Millipore 05-368 Curated Info
47	Inoue Y, Kitagawa M, Taya Y (2007) Phosphorylation of pRB at Ser612 by Chk1/2 leads to a complex between pRB and E2F-1 after DNA damage. EMBO J 26, 2083-93 17380128 Curated Info
48	Possemato A (2007) CST Curation Set: 2724; Year: 2007; Biosample/Treatment: cell line, K562/untreated; Disease: chronic myelogenous leukemia; SILAC: -; Specificities of Antibodies Used to Purify Peptides prior to LCMS: p[ST]P Antibodies Used to Purify Peptides prior to LCMS: Phospho-(Ser) CDKs Substrate Antibody Cat#: 2324, PTMScan(R) Phospho-CDK Substrate Motif (K/RS*PXK/R) Immunoaffinity Beads Cat#: 1981 Curated Info
49	Schmitz NM, Hirt A, Aebi M, Leibundgut K (2006) Limited redundancy in phosphorylation of retinoblastoma tumor suppressor protein by cyclin-dependent kinases in acute lymphoblastic leukemia. Am J Pathol 169, 1074-9 16936279 Curated Info
50	Schmitz NM, Leibundgut K, Hirt A (2005) CDK2 catalytic activity and loss of nuclear tethering of retinoblastoma protein in childhood acute lymphoblastic leukemia. Leukemia 19, 1783-7 16107892 Curated Info
51	Panigone S, et al. (2000) pRb and Cdk regulation by N-(4-hydroxyphenyl)retinamide. Oncogene 19, 4035-41 10962560 Curated Info
52	Knudsen ES, Wang JY (1997) Dual mechanisms for the inhibition of E2F binding to RB by cyclin-dependent kinase-mediated RB phosphorylation. Mol Cell Biol 17, 5771-83 9315635 Curated Info