JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2006 Vol.7 No.5 P.373-376

http://doi.org/10.1631/jzus.2006.B0373

Functional analysis of helicase and three tandem HRDC domains of RecQ in Deinococcus radiodurans

Author(s): Huang Li-fen, Hua Xiao-ting, Lu Hui-ming, Gao Guan-jun, Tian Bing, Shen Bing-hui, Hua Yue-jin
Affiliation(s): Institute of Nuclear-Agricultural Science, Zhejiang University, Hangzhou 310029, China; more
Corresponding email(s): yjhua@zju.edu.cn
Key Words: RecQ, Deinococcus radiodurans, Helicase, HRDC

Share this article to： More <<< Previous Article \|Next Article >>>

Abstract: recQ is a highly conserved helicase necessary for maintaining genome stability in all organisms. Genome comparison showed that a homologue of recQ in Deinococcus radiodurans designated as DR1289 is a member of recQ family with unusual domain arrangement: a helicase domain, an recQ C-terminal domain, and surprisingly three HRDC domain repeats, whose function, however, remains obscure currently. Using an insertion deletion, we discovered that the DRrecQ mutation causes an increase in gamma radiation, hydroxyurea and mitomycine C and UV sensitivity. Using the shuttle plasmid pRADK, we complemented various domains of the D. radiodurans recQ (DRrecQ) to the mutant in vivo. Results suggested that both the helicase and helicase-and-RNase-D-C-terminal (HRDC) domains are essential for complementing several phenotypes. The complementation and biochemical function of DRrecQ variants with different domains truncated in vitro suggested that both the helicase and three HRDC domains are necessary for recQ functions in D. radiodurans, while three HRDC domains have a synergistic effect on the whole function. Our finding leads to the hypothesis that the RecF recombination pathway is likely a primary path of double strand break repair in this well-known radioresistant organism.

Reference

[1] Daly, M.J., Ouyang, L., Fuchs, P., Minton, K.W., 1994. In vivo damage and recA-dependent repair of plasmid and chromosomal DNA in the radiation-resistant bacterium Deinococcus radiodurans. J. Bacteriol., 176(12):3508-3517.

[2] Eggington, J.M., Haruta, N., Wood, E.A., Cox, M.M., 2004. The single-stranded DNA-binding protein of Deinococcus radiodurans. BMC Microbiol., 4(1):2.

[3] Hishida, T., Han, Y.W., Shibata, T., Kubota, Y., Ishino, Y., Iwasaki, H., Shinagawa, H., 2004. Role of the Escherichia coli RecQ DNA helicase in SOS signaling and genome stabilization at stalled replication forks. Genes Dev., 18(15):1886-1897.

[4] Huber, M.D., Lee, D.C., Maizels, N., 2002. G4 DNA unwinding by BLM and Sgs1p: substrate specificity and substrate-specific inhibition. Nucleic Acids Res., 30(18):3954-3961.

[5] Khakhar, R.R., Cobb, J.A., Bjergbaek, L., Hickson, I.D., Gasser, S.M., 2003. RecQ helicases: multiple roles in genome maintenance. Trends. Cell Biol., 13(9):493-501.

[6] Makarova, K.S., Aravind, L., Wolf, Y.I., Tatusov, R.L., Minton, K.W., Koonin, E.V., Daly, M.J., 2001. Genome of the extremely radiation-resistant bacterium Deinococcus radiodurans viewed from the perspective of comparative genomics. Microbiol. Mol. Biol. Rev., 65(1):44-79.

[7] Minton, K.W., 1994. DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans. Mol. Microbiol., 13(1):9-15.

[8] Morozov, V., Mushegian, A.R., Koonin, E.V., Bork, P., 1997. A putative nucleic acid-binding domain in Bloom’s and Werner’s syndrome helicases. Trends Biochem. Sci., 22(11):417-418.

[9] Nakayama, M., Kawasaki, K., Matsumoto, K., Shibata, T., 2004. The possible roles of the DNA helicase and C-terminal domains in RECQ5/QE: complementation study in yeast. DNA Repair, 3(4):369-378.

[10] White, O., Eisen, J.A., Heidelberg, J.F., Hickey, E.K., Peterson, J.D., Dodson, R.J., Haft, D.H., Gwinn, M.L., Nelson, W.C., Richardson, D.L., et al., 1999. Genome sequence of the radioresistant bacterium Deinococcus radiodurans R1. Science, 286(5444):1571-1577.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Similar articles

- Go to

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference