Abstract
DNA polymerase eta carries out translesion synthesis past UV photoproducts and is deficient in xeroderma pigmentosum (XP) variants. We report that poleta is mostly localized uniformly in the nucleus but is associated with replication foci during S phase. Following treatment of cells with UV irradiation or carcinogens, it accumulates at replication foci stalled at DNA damage. The C-terminal third of poleta is not required for polymerase activity. However, the C-terminal 70 aa are needed for nuclear localization and a further 50 aa for relocalization into foci. Poleta truncations lacking these domains fail to correct the defects in XP-variant cells. Furthermore, we have identified mutations in two XP variant patients that leave the polymerase motifs intact but cause loss of the localization domains.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cell Line, Transformed
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Cell Nucleus / enzymology
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DNA / metabolism
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DNA / radiation effects
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DNA Damage
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DNA Primers / genetics
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DNA Repair
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DNA-Binding Proteins / metabolism
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DNA-Directed DNA Polymerase / chemistry
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DNA-Directed DNA Polymerase / genetics*
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DNA-Directed DNA Polymerase / metabolism*
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Genetic Variation
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Green Fluorescent Proteins
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Humans
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Luminescent Proteins / genetics
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Luminescent Proteins / metabolism
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Molecular Sequence Data
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Mutation
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Proliferating Cell Nuclear Antigen / metabolism
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Protein Structure, Tertiary
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Rad51 Recombinase
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Sequence Deletion
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Sequence Homology, Amino Acid
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Ultraviolet Rays / adverse effects
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Xeroderma Pigmentosum / enzymology*
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Xeroderma Pigmentosum / genetics*
Substances
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DNA Primers
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DNA-Binding Proteins
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Luminescent Proteins
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Proliferating Cell Nuclear Antigen
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Recombinant Fusion Proteins
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Green Fluorescent Proteins
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DNA
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RAD51 protein, human
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Rad51 Recombinase
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DNA-Directed DNA Polymerase
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Rad30 protein