Genetics, Vol. 166, 661-668, February 2004, Copyright © 2004

Developing a Genetic System in Deinococcus radiodurans for Analyzing Mutations

Mandy Kima, Erika Wolffa, Tiffany Huanga, Lilit Garibyana, Ashlee M. Earlb, John R. Battistab, and Jeffrey H. Millera
a Department of Microbiology, Immunology and Molecular Genetics and the Molecular Biology Institute, University of California, Los Angeles, California 90095
b Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803

Corresponding author: Jeffrey H. Miller, Immunology and Molecular Genetics and the Molecular Biology Institute, University of California, 609 Charles E. Young Dr. #1602, Los Angeles, CA 90095., jhmiller{at}mbi.ucla.edu (E-mail)

Communicating editor: S. T. LOVETT

We have applied a genetic system for analyzing mutations in Escherichia coli to Deinococcus radiodurans, an extremeophile with an astonishingly high resistance to UV- and ionizing-radiation-induced mutagenesis. Taking advantage of the conservation of the ß-subunit of RNA polymerase among most prokaryotes, we derived again in D. radiodurans the rpoB/Rifr system that we developed in E. coli to monitor base substitutions, defining 33 base change substitutions at 22 different base pairs. We sequenced >250 mutations leading to Rifr in D. radiodurans derived spontaneously in wild-type and uvrD (mismatch-repair-deficient) backgrounds and after treatment with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and 5-azacytidine (5AZ). The specificities of NTG and 5AZ in D. radiodurans are the same as those found for E. coli and other organisms. There are prominent base substitution hotspots in rpoB in both D. radiodurans and E. coli. In several cases these are at different points in each organism, even though the DNA sequences surrounding the hotspots and their corresponding sites are very similar in both D. radiodurans and E. coli. In one case the hotspots occur at the same site in both organisms.




This article has been cited by other articles:


Home page
 J. Bacteriol.Home page
A. M. Marcobal, D. A. Sela, Y. I. Wolf, K. S. Makarova, and D. A. Mills
Role of Hypermutability in the Evolution of the Genus Oenococcus
J. Bacteriol., January 15, 2008; 190(2): 564 - 570.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
K. Zeibell, S. Aguila, V. Yan Shi, A. Chan, H. Yang, and J. H. Miller
Mutagenesis and Repair in Bacillus anthracis: the Effect of Mutators
J. Bacteriol., March 15, 2007; 189(6): 2331 - 2338.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
R. S. Galhardo, R. P. Rocha, M. V. Marques, and C. F. M. Menck
An SOS-regulated operon involved in damage-inducible mutagenesis in Caulobacter crescentus
Nucleic Acids Res., May 10, 2005; 33(8): 2603 - 2614.
[Abstract] [Full Text] [PDF]