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Assessment of CDK4 as a serine kinase phosphorylating OGG1

ABSTRACT: The early studies showed the serine/threonine of OGG1 was phosphorylated by CDK4 in vitro, in our study, LC-MS/MS was performed, and Byonic engine was used to identify the precise site to be phosphorylated by CDK4.The present study discovered a high basal level of CDK4-mediated phosphorylation of OGG1 at S326, and this modification undergoes decline to resume a higher affinity with DNA for facilitating the role of OGG1 in gene transcription.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Lung

SUBMITTER: Jinling Han

LAB HEAD: Professor Xueqing Ba

PROVIDER: PXD059565 | Pride | 2025-04-18

REPOSITORIES: Pride

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			Action	DRS
	BPI_32365_ECOLI_HJ11104.mgf	Mgf
	BPI_32365_ECOLI_HJ11104.mzid.gz	Mzid
	OGG1Phosphorylation-protein-Rep2.msf	Msf
	OGG1_phosphorylation_0_Rep2.raw	Raw
	checksum.txt	Txt

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Project description:The retinoblastoma tumor suppressor protein (Rb) regulates early G1 phase checkpoints, including the DNA damage response, as well as cell cycle exit and differentiation. The widely accepted model of G1 cell cycle progression proposes that cyclin D:Cdk4/6 partially inactivates the Rb tumor suppressor during early G1 phase by progressive multi-phosphorylation, termed hypo-phosphorylation, resulting in release of E2F transcription factors. However, this model remains largely unproven biochemically and the biologically active form(s) of Rb remains unknown. Here we find that Rb is un-phosphorylated in G0 cells and becomes exclusively mono-phosphorylated throughout all of early G1 phase by cyclin D:Cdk4/6. Early G1 phase mono-phosphorylated Rb is composed of 14 independent isoforms that are all targeted by the E1a oncoprotein, but each shows a preferential binding pattern to specific E2F1-4 transcription factors. At the late G1 Restriction Point, cyclin E:Cdk2 inactivates Rb by a quantum hyper-phosphorylation (>12 phosphates/Rb). Cells undergoing a DNA damage response activate cyclin D:Cdk4/6 to generate mono-phosphorylated Rb that regulates global transcription. In contrast, a non-phosphorylatable ?Cdk-Rb allele was non-functional for regulating a DNA damage response, but functional for driving cell cycle exit and differentiation during myogenesis. These observations fundamentally change our understanding of G1 cell cycle progression and show that there is no progressive multi-phosphorylation or hypo-phosphorylation inactivation of Rb during early G1 phase by cyclin D:Cdk4/6. Instead, cyclin D:Cdk4/6 generates functionally active, mono-phosphorylated Rb that is the only Rb isoform present in cells during early G1 phase. Global transcriptional analysis of murine embryonic fibroblasts (MEFs) with conditional deletion of the endogenous RB gene by treatment with cell permeable TAT-Cre. Comparison to unaltered MEFs and MEFs with physiological level of exogenous wildtype or phospho-mutant RB expressed at time of RB gene deletion.

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Assessment of CDK4 as a serine kinase phosphorylating OGG1

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