Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana

Journal Article · Fri Jan 01 00:00:00 EST 2010 · Journal of Nucleic Acids

DOI:https://doi.org/10.4061/2010/701472· OSTI ID:1198461

Kashiwagi, Sayo ^[1]; Kuraoka, Isao ^[1]; Fujiwara, Yoshie ^[1]; Hitomi, Kenichi ^[2]; Cheng, Quen J. ^[3]; Fuss, Jill O. ^[4]; Shin, David S. ^[5]; Masutani, Chikahide ^[6]; Tainer, John A. ^[5]; Hanaoka, Fumio ^[7]; Iwai, Shigenori ^[1]

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1. Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan
2. Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan, Department of Molecular Biology and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA, Life Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
3. Life Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA, University of California San Diego, 9500 Gilman Drive no. 0613C, La Jolla, CA 92093, USA
4. Life Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
5. Department of Molecular Biology and The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA, Life Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
6. Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan
7. Faculty of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588, Japan

Human DNA polymerase $\eta$ (HsPol $\eta$ ) plays an important role in translesion synthesis (TLS), which allows for replication past DNA damage such as UV-induced cis-syn cyclobutane pyrimidine dimers (CPDs). Here, we characterized ApPol $\eta$ from the thermophilic worm Alvinella pompejana , which inhabits deep-sea hydrothermal vent chimneys. ApPol $\eta$ shares sequence homology with HsPol $\eta$ and contains domains for binding ubiquitin and proliferating cell nuclear antigen. Sun-induced UV does not penetrate Alvinella's environment; however, this novel DNA polymerase catalyzed efficient and accurate TLS past CPD, as well as 7,8-dihydro-8-oxoguanine and isomers of thymine glycol induced by reactive oxygen species. In addition, we found that ApPol $\eta$ is more thermostable than HsPol $\eta$ , as expected from its habitat temperature. Moreover, the activity of this enzyme was retained in the presence of a higher concentration of organic solvents. Therefore, ApPol $\eta$ provides a robust, human-like Pol $\eta$ that is more active after exposure to high temperatures and organic solvents.

• Sponsoring Organization: USDOE
• Grant/Contract Number: FG0207ER64326
• OSTI ID: 1198461
• Journal Information: Journal of Nucleic Acids, Journal Name: Journal of Nucleic Acids Vol. 2010; ISSN 2090-021X
• Publisher: Hindawi Publishing Corporation
• Country of Publication: Country unknown/Code not available
• Language: English