Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana
Abstract
Human DNA polymerase (HsPol ) 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 from the thermophilic worm Alvinella pompejana , which inhabits deep-sea hydrothermal vent chimneys. ApPol shares sequence homology with HsPol 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 is more thermostable than HsPol , 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 provides a robust, human-like Pol that is more active after exposure to high temperatures and organic solvents.
- Authors:
-
- Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka, Osaka 560-8531, Japan
- 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
- 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
- Life Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
- 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
- Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan
- Faculty of Science, Gakushuin University, 1-5-1 Mejiro, Toshima-ku, Tokyo 171-8588, Japan
- Publication Date:
- Sponsoring Org.:
- USDOE
- OSTI Identifier:
- 1198461
- Grant/Contract Number:
- FG0207ER64326
- Resource Type:
- Journal Article: Published Article
- Journal Name:
- Journal of Nucleic Acids
- Additional Journal Information:
- Journal Name: Journal of Nucleic Acids Journal Volume: 2010; Journal ID: ISSN 2090-021X
- Publisher:
- Hindawi Publishing Corporation
- Country of Publication:
- Country unknown/Code not available
- Language:
- English
Citation Formats
Kashiwagi, Sayo, Kuraoka, Isao, Fujiwara, Yoshie, Hitomi, Kenichi, Cheng, Quen J., Fuss, Jill O., Shin, David S., Masutani, Chikahide, Tainer, John A., Hanaoka, Fumio, and Iwai, Shigenori. Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana. Country unknown/Code not available: N. p., 2010.
Web. doi:10.4061/2010/701472.
Kashiwagi, Sayo, Kuraoka, Isao, Fujiwara, Yoshie, Hitomi, Kenichi, Cheng, Quen J., Fuss, Jill O., Shin, David S., Masutani, Chikahide, Tainer, John A., Hanaoka, Fumio, & Iwai, Shigenori. Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana. Country unknown/Code not available. https://doi.org/10.4061/2010/701472
Kashiwagi, Sayo, Kuraoka, Isao, Fujiwara, Yoshie, Hitomi, Kenichi, Cheng, Quen J., Fuss, Jill O., Shin, David S., Masutani, Chikahide, Tainer, John A., Hanaoka, Fumio, and Iwai, Shigenori. 2010.
"Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana". Country unknown/Code not available. https://doi.org/10.4061/2010/701472.
@article{osti_1198461,
title = {Characterization of a Y-Family DNA Polymerase eta from the Eukaryotic Thermophile Alvinella pompejana},
author = {Kashiwagi, Sayo and Kuraoka, Isao and Fujiwara, Yoshie and Hitomi, Kenichi and Cheng, Quen J. and Fuss, Jill O. and Shin, David S. and Masutani, Chikahide and Tainer, John A. and Hanaoka, Fumio and Iwai, Shigenori},
abstractNote = {Human DNA polymerase η (HsPol η ) 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 η from the thermophilic worm Alvinella pompejana , which inhabits deep-sea hydrothermal vent chimneys. ApPol η shares sequence homology with HsPol η 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 η is more thermostable than HsPol η , 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 η provides a robust, human-like Pol η that is more active after exposure to high temperatures and organic solvents.},
doi = {10.4061/2010/701472},
url = {https://www.osti.gov/biblio/1198461},
journal = {Journal of Nucleic Acids},
issn = {2090-021X},
number = ,
volume = 2010,
place = {Country unknown/Code not available},
year = {Fri Jan 01 00:00:00 EST 2010},
month = {Fri Jan 01 00:00:00 EST 2010}
}