TitleArabidopsis thaliana AtPOLK encodes a DinB-like DNA polymerase that extends mispaired primer termini and is highly expressed in a variety of tissues.
Publication TypeJournal Article
Year of Publication2004
AuthorsGarcía-Ortiz MVictoria, Ariza RR, Hoffman PD, Hays JB, Roldán-Arjona T
JournalPlant J
Date Published2004 Jul
Alternative Splicing, Amino Acid Sequence, Arabidopsis, Arabidopsis Proteins, DNA-Directed DNA Polymerase, Gene Expression, Gene Library, Molecular Sequence Data, Phylogeny, Plants, Genetically Modified, Recombinant Fusion Proteins, Sequence Homology, Amino Acid

Cell survival after DNA damage depends on specialized DNA polymerases able to perform DNA synthesis on imperfect templates. Most of these enzymes belong to the recently discovered Y-family of DNA polymerases, none of which has been previously described in plants. We report here the isolation, functional characterization and expression analysis of a plant representative of the Y-family. This polymerase, which we have termed AtPolkappa, is a homolog of Escherichia coli pol IV and human pol kappa, and thus belongs to the DinB subfamily. We purified AtPolkappa and found a template-directed DNA polymerase, endowed with limited processivity that is able to extend primer-terminal mispairs. The activity and processivity of AtPolkappa are enhanced markedly upon deletion of 193 amino acids (aa) from its carboxy (C)-terminal domain. Loss of this region also affects the nucleotide selectivity of the enzyme, leading to the incorporation of both dCTP and dTTP opposite A in the template. We detected three cDNA forms, which result from the alternative splicing of AtPOLK mRNA and have distinct patterns of expression in different plant organs. Histochemical localization of beta-glucuronidase (GUS) activity in transgenic plants revealed that the AtPOLK promoter is active in endoreduplicating cells, suggesting a possible role during consecutive DNA replication cycles in the absence of mitosis.

Alternate JournalPlant J.
PubMed ID15200644