Characterization of a DNA damage-inducible membrane protein kinase from Deinococcus radiodurans and its role in bacterial radioresistance and DNA strand break repair

Differential hybridization has been used to identify genes in Saccharomyces cerevisiae displaying increased transcript levels after treatment of cells with UV irradiation or with the mutagen/carcinogen 4-nitroquinoline-1-oxide (NQO). We describe the isolation and characterization of four DNA damage responsive genes obtained from screening ca. 9,000 yeast genomic clones. Two of these clones, lambda 78A and pBR178C, contain repetitive elements in the yeast genome as shown by Southern hybridization analysis. Although the genomic hybridization pattern is distinct for each of these two clones, both of these sequences hybridize to large polyadenylated transcripts ca. 5 kilobases in length. Two other DNA damage responsive sequences, pBRA2 and pBR3016B, are single-copy genes and hybridize to 0.5- and 3.2-kilobase transcripts, respectively. Kinetic analysis of the 0.5-kilobase transcript homologous to pBRA2 indicates that the level of this RNA increases more than 15-fold within 20 min after exposure to 4-nitroquinoline-1-oxide. Moreover, the level of this transcript is significantly elevated in cells containing the rad52-1 mutation which are deficient in DNA strand break repair and gene conversion. These results provide some of the first evidence that DNA damage stimulates transcription of specific genes in eucaryotic cells.

Download Full-text

The SbcCD complex of Deinococcus radiodurans contributes to radioresistance and DNA strand break repair in vivo and exhibits Mre11–Rad50 type activity in vitro

DNA Repair ◽

10.1016/j.dnarep.2010.01.012 ◽

2010 ◽

Vol 9 (5) ◽

pp. 488-494 ◽

Cited By ~ 15

Author(s):

Vidya A. Kamble ◽

Hari S. Misra

Keyword(s):

Deinococcus Radiodurans ◽

Strand Break ◽

Dna Strand Break ◽

Type Activity ◽

Break Repair ◽

Dna Strand

Download Full-text

Involvement of a periplasmic protein kinase in DNA strand break repair and homologous recombination in Escherichia coli

Molecular Microbiology ◽

10.1111/j.1365-2958.2007.05919.x ◽

2007 ◽

Vol 65 (6) ◽

pp. 1595-1595

Author(s):

Nivedita P. Khairnar ◽

Vidya A. Kamble ◽

Suhas H. Mangoli ◽

Shree K. Apte ◽

Hari S. Misra

Keyword(s):

Escherichia Coli ◽

Protein Kinase ◽

Homologous Recombination ◽

Strand Break ◽

Periplasmic Protein ◽

Dna Strand Break ◽

Break Repair ◽

Dna Strand

Download Full-text

PprA Protein Inhibits DNA Strand Exchange and ATP Hydrolysis of Deinococcus RecA and Regulates the Recombination in Gamma-Irradiated Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.636178 ◽

2021 ◽

Vol 9 ◽

Author(s):

Yogendra Singh Rajpurohit ◽

Dhirendra Kumar Sharma ◽

Hari S. Misra

Keyword(s):

Deinococcus Radiodurans ◽

Strand Break ◽

Strand Exchange ◽

Binding Property ◽

Functional Interaction ◽

Dna Strand Break ◽

Dna Strand Exchange ◽

Break Repair ◽

Dna Strand ◽

Gamma Irradiated

DrRecA and PprA proteins function are crucial for the extraordinary resistance to γ-radiation and DNA strand break repair in Deinococcus radiodurans. DrRecA mediated homologous recombination help in DNA strand break repair and cell survival, while the PprA protein confers radio-resistance via its roles in DNA repair, genome maintenance, and cell division. Genetically recA and pprA genes interact and constitute an epistatic group however, the mechanism underlying their functional interaction is not clear. Here, we showed the physical and functional interaction of DrRecA and PprA protein both in solution and inside the cells. The absence of the pprA gene increases the recombination frequency in gamma-irradiated D. radiodurans cells and genomic instability in cells growing under normal conditions. PprA negatively regulates the DrRecA functions by inhibiting DrRecA mediated DNA strand exchange and ATPase function in vitro. Furthermore, it is shown that the inhibitory effect of PprA on DrRecA catalyzed DNA strand exchange was not due to sequestration of homologous dsDNA and was dependent on PprA oligomerization and DNA binding property. Together, results suggest that PprA is a new member of recombination mediator proteins (RMPs), and able to regulate the DrRecA function in γ-irradiated cells by protecting the D. radiodurans genome from hyper-recombination and associated negative effects.

Download Full-text

Inactivation of Cdc2 increases the level of apoptosis induced by DNA damage

Journal of Cell Science ◽

10.1242/jcs.108.8.2897 ◽

1995 ◽

Vol 108 (8) ◽

pp. 2897-2904 ◽

Cited By ~ 1

Author(s):

W. Ongkeko ◽

D.J. Ferguson ◽

A.L. Harris ◽

C. Norbury

Keyword(s):

Dna Damage ◽

Cell Death ◽

Protein Kinase ◽

Apoptotic Cell ◽

Mutant Cell ◽

Strand Break ◽

Temperature Sensitive ◽

Dna Strand Break ◽

Dna Strand ◽

Inhibitor Compounds

A number of lines of evidence have suggested a possible involvement of the mitosis-promoting protein kinase Cdc2 in the process of apoptotic cell death, and one recent study concluded that premature activation of Cdc2 is required for apoptosis. Here we have used a temperature-sensitive murine Cdc2 mutant cell line and Cdc2 inhibitor compounds to study the effect of inhibition of this protein kinase on apoptosis induced by DNA-damaging drugs. Inhibition of Cdc2 activity before or during exposure to DNA strand break-inducing drugs had the effect of increasing the level of subsequent apoptosis, as assessed by electron microscopy and flow cytometry. We conclude that, far from being required for cell death, a form of mammalian Cdc2 suppresses apoptosis induced by DNA damage. This form of Cdc2 appears to be active in G2-arrested cells and is therefore presumably distinct from the mitosis-promoting Cdc2-cyclin B heterodimer.

Download Full-text