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Identification of new genes contributing to the extreme radioresistance of Deinococcus radiodurans using a Tn5-based transposon mutant library.

Dulermo R, Onodera T, Coste G, Passot F, Dutertre M, Porteron M, Confalonieri F, Sommer S, Pasternak C - PLoS ONE (2015)

Bottom Line: Here, we have developed an extremely efficient in vivo Tn5-based mutagenesis procedure to construct a Deinococcus radiodurans insertion mutant library subsequently screened for sensitivity to genotoxic agents such as γ and UV radiations or mitomycin C.Interestingly, preliminary characterization of previously undescribed radiosensitive mutants suggests the contribution of cyclic di-AMP signaling in the recovery of D. radiodurans cells from genotoxic stresses, probably by modulating several pathways involved in the overall cell response.Our analyses also point out a new transcriptional regulator belonging to the GntR family, encoded by DR0265, and a predicted RNase belonging to the newly described Y family, both contributing to the extreme radioresistance of D. radiodurans.

View Article: PubMed Central - PubMed

Affiliation: Univ. Paris-Sud, Institute for Integrative Biology of the Cell (I2BC), Université Paris Saclay, CEA, CNRS, Orsay, France.

ABSTRACT
Here, we have developed an extremely efficient in vivo Tn5-based mutagenesis procedure to construct a Deinococcus radiodurans insertion mutant library subsequently screened for sensitivity to genotoxic agents such as γ and UV radiations or mitomycin C. The genes inactivated in radiosensitive mutants belong to various functional categories, including DNA repair functions, stress responses, signal transduction, membrane transport, several metabolic pathways, and genes of unknown function. Interestingly, preliminary characterization of previously undescribed radiosensitive mutants suggests the contribution of cyclic di-AMP signaling in the recovery of D. radiodurans cells from genotoxic stresses, probably by modulating several pathways involved in the overall cell response. Our analyses also point out a new transcriptional regulator belonging to the GntR family, encoded by DR0265, and a predicted RNase belonging to the newly described Y family, both contributing to the extreme radioresistance of D. radiodurans. Altogether, this work has revealed new cell responses involved either directly or indirectly in repair of various cell damage and confirmed that D. radiodurans extreme radiation resistance is determined by a multiplicity of pathways acting as a complex network.

No MeSH data available.


Related in: MedlinePlus

Measurement of sensitivity to H2O2 stress by disc inhibition assay (procedure described in Materials and Methods).(A) Phenotype of the wild-type (resistant), the katA mutant (highly sensitive), and the DR0265 mutant (middle sensitive) are shown. (B) The mutants are classified into four categories depending on the diameter (in mm) of growth inhibition area as indicated.
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pone.0124358.g002: Measurement of sensitivity to H2O2 stress by disc inhibition assay (procedure described in Materials and Methods).(A) Phenotype of the wild-type (resistant), the katA mutant (highly sensitive), and the DR0265 mutant (middle sensitive) are shown. (B) The mutants are classified into four categories depending on the diameter (in mm) of growth inhibition area as indicated.

Mentions: c by using the disc inhibition assay as described in Materials and Methods and Fig 2 (for the inhibition diameters corresponding to the different levels of sensitivity).


Identification of new genes contributing to the extreme radioresistance of Deinococcus radiodurans using a Tn5-based transposon mutant library.

Dulermo R, Onodera T, Coste G, Passot F, Dutertre M, Porteron M, Confalonieri F, Sommer S, Pasternak C - PLoS ONE (2015)

Measurement of sensitivity to H2O2 stress by disc inhibition assay (procedure described in Materials and Methods).(A) Phenotype of the wild-type (resistant), the katA mutant (highly sensitive), and the DR0265 mutant (middle sensitive) are shown. (B) The mutants are classified into four categories depending on the diameter (in mm) of growth inhibition area as indicated.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401554&req=5

pone.0124358.g002: Measurement of sensitivity to H2O2 stress by disc inhibition assay (procedure described in Materials and Methods).(A) Phenotype of the wild-type (resistant), the katA mutant (highly sensitive), and the DR0265 mutant (middle sensitive) are shown. (B) The mutants are classified into four categories depending on the diameter (in mm) of growth inhibition area as indicated.
Mentions: c by using the disc inhibition assay as described in Materials and Methods and Fig 2 (for the inhibition diameters corresponding to the different levels of sensitivity).

Bottom Line: Here, we have developed an extremely efficient in vivo Tn5-based mutagenesis procedure to construct a Deinococcus radiodurans insertion mutant library subsequently screened for sensitivity to genotoxic agents such as γ and UV radiations or mitomycin C.Interestingly, preliminary characterization of previously undescribed radiosensitive mutants suggests the contribution of cyclic di-AMP signaling in the recovery of D. radiodurans cells from genotoxic stresses, probably by modulating several pathways involved in the overall cell response.Our analyses also point out a new transcriptional regulator belonging to the GntR family, encoded by DR0265, and a predicted RNase belonging to the newly described Y family, both contributing to the extreme radioresistance of D. radiodurans.

View Article: PubMed Central - PubMed

Affiliation: Univ. Paris-Sud, Institute for Integrative Biology of the Cell (I2BC), Université Paris Saclay, CEA, CNRS, Orsay, France.

ABSTRACT
Here, we have developed an extremely efficient in vivo Tn5-based mutagenesis procedure to construct a Deinococcus radiodurans insertion mutant library subsequently screened for sensitivity to genotoxic agents such as γ and UV radiations or mitomycin C. The genes inactivated in radiosensitive mutants belong to various functional categories, including DNA repair functions, stress responses, signal transduction, membrane transport, several metabolic pathways, and genes of unknown function. Interestingly, preliminary characterization of previously undescribed radiosensitive mutants suggests the contribution of cyclic di-AMP signaling in the recovery of D. radiodurans cells from genotoxic stresses, probably by modulating several pathways involved in the overall cell response. Our analyses also point out a new transcriptional regulator belonging to the GntR family, encoded by DR0265, and a predicted RNase belonging to the newly described Y family, both contributing to the extreme radioresistance of D. radiodurans. Altogether, this work has revealed new cell responses involved either directly or indirectly in repair of various cell damage and confirmed that D. radiodurans extreme radiation resistance is determined by a multiplicity of pathways acting as a complex network.

No MeSH data available.


Related in: MedlinePlus