caa a22 4500 397557981 CHVBK 20180308164808.0 cr unu---uuuuu 161202e199610 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021477 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021477 Replication Protein-A Mediates the Association of Calf Thymus DNA Polymerase α-DNA Primase Complex with Guanine-Rich DNA Sequence [Elektronische Daten] [Motoshi Suzuki, Keiko Tamiya-Koizumi, Masaharu Takemura, Keigo Furuta, Shunji Izuta, Ericka Savoysky, Ayuko Miura, Shonen Yoshida] We have shown that calf thymus DNA polymerase α-DNA primase complex (polα-primase) preferentially binds to pyrimidine-rich sequences and initiates RNA primer synthesis [Suzuki, M. et at. (1993) Biochemistry 32, 12782-12792]. Here we tested the association of polα-primase with a guanine-rich DNA fragment (SVG, 30-mer) containing in vivo initiation sites of simian virus 40 DNA replication. While pyrimidine-rich fragment (CTPPS 1, 30-mer), that is a preferred sequence for calf thymus DNA primase, was well co-precipitated with polα-primase using anti-polα antibody, SVG was hardly precipitated under the same conditions. Competition studies in either gel-retardation assay or during de novo DNA synthesis by polα-primase demonstrated that the interaction of polα-primase with SVG was much weaker than that with CTPPS 1. On the other hand, replication protein-A (RP-A) could bind SVG, although less efficiently than CTPPS 1. After preincubation with RP-A, SVG could bind polα-primase that was immobilized on Sepharose beads. The simian virus 40 large T antigen also enhanced association of SVG to polα-primase, while Escherichia coli single-stranded DNA-binding protein did not. However, polα-primase, bound to SVG in the presence of RP-A, failed to synthesize RNA primers. When SVG was extended 10 nucleotldes at its 5′ -end, polα-primase synthesized trace amounts of RNA primers, and this activity was stimulated more than 10-fold by adding RP-A. These results suggest a new role for RP-A, i.e., as a molecular tether that allows polα-primase to bind guanine-rich regions of DNA in order to initiate RNA primer synthesis. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence DNA polymerase α-primase nationallicence DNA replication nationallicence replication protein A nationallicence Suzuki Motoshi Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut Tamiya-Koizumi Keiko Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut Takemura Masaharu Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut Furuta Keigo Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut Izuta Shunji Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut Savoysky Ericka Chugai Pharmaceutical Co., Ltd, Fuji-Gotemba Research Labs 135 Komakado 1-chome, Gotemba, Shizuoka 412 aut Miura Ayuko Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut Yoshida Shonen Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut The Journal of Biochemistry Oxford University Press 120/4(1996-10), 766-772 0021-924X 120:4<766 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021477 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021477 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Suzuki Motoshi Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Tamiya-Koizumi Keiko Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Takemura Masaharu Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Furuta Keigo Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Izuta Shunji Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Savoysky Ericka Chugai Pharmaceutical Co., Ltd, Fuji-Gotemba Research Labs 135 Komakado 1-chome, Gotemba, Shizuoka 412 aut NATIONALLICENCE 700 1- Miura Ayuko Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 700 1- Yoshida Shonen Laboratory of Cancer Cell Biology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine 65 Tsurumai-cho, Showa-ku, Nagoya 466 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/4(1996-10), 766-772 0021-924X 120:4<766 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford