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kota, S.

Publications and source records attributed to kota, S..

3 recordsLinked to original sources

In vivo guanine quadruplex structure dynamics and role in genome maintenance in Deinococcus radiodurans

Guanine quadruplex (G4) structures are secondary structures formed in nucleic acids that are rich in guanine bases. G4 structures regulate various cellular processes both in eukaryotes and prokaryotes. Deinococcus radiodurans, an extremophile show sensitivity to ionizing radiation in the presence of G4 binding ligands during the post-irradiation recovery period (PIR). Putative G4 motifs positioned at different locations on the genes fold into different topologies in vitro in the presence of monovalent cations while divalent cation Mg+2 supported the stable G4 formation but Mn+2 destabilizes the G4 structures. Thioflavin T and anti-DNA quadruplex antibodies detected the in vivo formation and dynamics of G4 structure in response to various DNA damaging agents, more in the presence of gamma radiation treatment. G4 binding drugs during PIR arrested the G4 structure dynamics and delayed the DNA repair process. Further, the absence of G4 helicase, RecQ resulted in the accumulation of more G4 structures and genome instability. All these results indicate that the guanine quadruplex structure formation increases in response to cellular stress and G4s are crucial for stable genome maintenance in Deinococcus radiodurans.

microbiology↗

DivIVA phosphorylation at threonine affects its dynamics and cell cycle in Deinococcus radiodurans.

RqkA, a {gamma} radiation responsive Ser/Thr quinoprotein kinase, is characterized for its role in radioresistance in Deinoocccus radiodurans. DivIVA is a cell division protein involved in determination of cell pole and division site in bacteria. RqkA phosphorylated cognate DivIVA (drDivIVA) at Threonine 19 (T19) residue located in its pole recognition motif. The phospho-mimetic replacement of T19 (T19E) functioned differently than phospho-ablative (T19A) and drDivIVA proteins. T19E-RFP expressing in wild type background showed arrest in dynamics of drDivIVA, and loss of interaction with genome segregation protein. divIVA is shown to be an essential gene in this bacterium. The allelic replacement of divIVA with T19E-RFP was not tolerated unless drDivIVA was expressed episomally while there was no effect of this replacement with T19A-RFP and drDivIVA-RFP. These results suggested that the T19 phosphorylation in drDivIVA by RqkA has affected in vivo functions of DivIVA that would render cell cycle arrest in this bacterium.

microbiology↗

Topoisomerase IB interacts with genome segregation proteins and is involved in multipartite genome maintenance in Deinococcus radiodurans

Deinococcus radiodurans, an extremophile, resistant to many abiotic stresses including ionizing radiation, has 2 type I topoisomerases (drTopo IA and drTopo IB) and one type II topoisomerase (DNA gyrase). The role of drTopo IB in guanine quadruplex DNA (G4 DNA) metabolism was shown in vitro. Here we report that D. radiodurans cells lacking drTopo IB ({Delta}topoIB) show sensitivity to G4 DNA binding drug (NMM) under normal growth conditions. The activity of G4 motif containing promoters like mutL and recQ was reduced in the presence of NMM in mutant cells. In mutant cells, the percentage of anucleated cells was more while the ploidy numbers of genome elements were less as compared to wild type. Protein-protein interaction studies showed that drTopo IB interacts with genome segregation and DNA replication initiation (DnaA) proteins. The typical patterns of cellular localization of GFP-PprA were affected in the mutant cells. Microscopic examination of D. radiodurans cells expressing drTopo IB-RFP showed its localization on nucleoid forming a streak parallel to the old division septum and perpendicular to newly formed septum. These results together suggest the role of drTopo IB in genome maintenance in this bacterium.

microbiology↗