Abstract
Numerous proteins, many essential for the DNA replication machinery, interact with proliferating cell nuclear antigen (PCNA) through the PCNA-interacting peptide (PIP) sequence called the PIP box. We have previously shown that the oxidative demethylase human AlkB homologue 2 (hABH2) colocalizes with PCNA in replication foci. In this study, we show that hABH2 interacts with a posttranslationally modified PCNA via a novel PCNA-interacting motif, which we term AlkB homologue 2 PCNA-interacting motif (APIM). We identify APIM in >200 other proteins involved in DNA maintenance, transcription, and cell cycle regulation, and verify a functional APIM in five of these. Expression of an APIM peptide increases the cellular sensitivity to several cytostatic agents not accounted for by perturbing only the hABH2-PCNA interaction. Thus, APIM is likely to mediate PCNA binding in many proteins involved in DNA repair and cell cycle control during genotoxic stress.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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AlkB Homolog 2, Alpha-Ketoglutarate-Dependent Dioxygenase
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Amino Acid Motifs / genetics*
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Animals
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DNA Repair
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DNA Repair Enzymes / genetics
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DNA Repair Enzymes / metabolism*
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DNA Topoisomerases, Type II / genetics
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DNA Topoisomerases, Type II / metabolism
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Dioxygenases / genetics
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Dioxygenases / metabolism*
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Fluorescence Resonance Energy Transfer
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HeLa Cells
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Humans
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Molecular Sequence Data
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Point Mutation
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Proliferating Cell Nuclear Antigen / genetics
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Proliferating Cell Nuclear Antigen / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Sequence Alignment
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Transcription Factors, TFII / genetics
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Transcription Factors, TFII / metabolism
Substances
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Proliferating Cell Nuclear Antigen
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Recombinant Fusion Proteins
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Transcription Factors, TFII
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Dioxygenases
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ALKBH2 protein, human
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AlkB Homolog 2, Alpha-Ketoglutarate-Dependent Dioxygenase
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DNA Topoisomerases, Type II
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DNA Repair Enzymes