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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
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.
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. [doi:10.1186/1471-2180-4-2]
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. [doi:10.1101/gad.1223804]
Huber, M.D., Lee, D.C., Maizels, N., 2002. G4 DNA unwinding by BLM and Sgslp: substrate specificity and substrate-specific inhibition. Nucleic Acids Res., 30(18):3954–3961. [doi:10.1093/nar/gkf530]
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. [doi:10.1016/S0962-8924(03)00171-5]
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 genomies. Microbiol. Mol. Biol. Rev., 65(1):44–79. [doi:10.1128/MMBR.65.1.44-79.2001]
Minton, K.W., 1994. DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans. Mol. Microbiol., 13(1):9–15.
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. [doi:10.1016/S0968-0004(97)01128-6]
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. [doi:10.1016/j.dnarep.2003.12.001]
White, O., Eisen, J.A., Heidelberg, J.F., Hiekey, 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. [doi:10.1126/science.286.5444.1571]
Author information
Authors and Affiliations
Corresponding author
Additional information
Project supported by the National Basic Research Program (973) of China (No. 2004CB19604), Distinguished Young Scientist of China (No. 30425038), and the National Natural Science Foundation of China (No. 30330020)
Note: This is a part of our work submitted to DNA Repair on November 30, 2005. We were informed by the editor for revision on January 9, 2006. During revision, a similar work entitled “Three HRDC domains differentially modulate Deinococcus radiodurans RecQ helicase biochemical activity” by Micheal P. Killoran and James L. Keck was submitted to J. Biol. Chem. on January 3, 2006 and online on March 9, 2006
Rights and permissions
About this article
Cite this article
Huang, Lf., Hua, Xt., LU, Hm. et al. Functional analysis of helicase and three tandem HRDC domains of RecQ in Deinococcus radiodurans . J. Zhejiang Univ. - Sci. B 7, 373–376 (2006). https://doi.org/10.1631/jzus.2006.B0373
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1631/jzus.2006.B0373