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Molecular cloning of the cDNA for the catalytic subunit of plant DNA polymerase α and its cell‐cycle dependent expression
Author(s) -
Yokoi Masayuki,
Ito Masaki,
Izumi Masako,
Miyazawa Hiroshi,
Nakai Hirokazu,
Hanaoka Fumio
Publication year - 1997
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1046/j.1365-2443.1997.1560354.x
Subject(s) - biology , cloning (programming) , complementary dna , dna polymerase , molecular cloning , protein subunit , microbiology and biotechnology , computational biology , dna , genetics , gene , computer science , programming language
Background: DNA polymerase α has been studied in considerable detail in yeast and animals. Genetic and biochemical analyses reveal that this enzyme is composed of a heterotetramer and is necessary for replicon initiation and primer synthesis in lagging strand synthesis. In spite of the fact that modes of DNA replication in plants seem to be similar to those in other eukaryotes, very little is known about the biochemical components that participate in DNA replication of plants, including DNA polymerases. Results: Using a 561‐base pair DNA fragment, obtained by polymerase chain reaction amplification from a rice cDNA library as a probe, we isolated and sequenced a cDNA homologous to the cDNA for the catalytic subunit of rice DNA polymerase α. The encoded polypeptide has extensive homology with the catalytic subunit of DNA polymerase α from several species. Furthermore, when the cDNA was expressed in eukaryotic transcription/translation systems, the protein products showed DNA polymerase activity which was inhibited by a monoclonal antibody specific for DNA polymerase α. Using RNA gel blot analysis, we found that the levels of mRNA of the catalytic subunit of this enzyme is regulated during the cell‐cycle in plant cells. Conclusion: This is the first report which describes the cDNA cloning of plant DNA polymerase. We conclude that the principal features of the DNA polymerase α catalytic subunit are conserved in plants.