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Title:	The nucleoid protein Dps binds genomic DNA of Escherichia coli in a non-random manner
Author:	Antipov, Sergey S.; Tutukina, Maria; Preobrazhenskaya, Elena V.; Kondrashov, Fyodor A., 1979-; Patrushev, Maksim Vladimirovich; Toshchakov, Stepan; Dominova, Irina N.; Shvyreva, Uliana S.; Vrublevskaya, Veronika V.; Morenkov, Oleg S.; Sukharicheva, Natalya A.; Panyukov, Valeriy V.; Ozoline, Olga N.
Abstract:	Dps is a multifunctional homododecameric protein that oxidizes Fe2+ ions accumulating them in the form of Fe2O3 within its protein cavity, interacts with DNA tightly condensing bacterial nucleoid upon starvation and performs some other functions. During the last two decades from discovery of this protein, its ferroxidase activity became rather well studied, but the mechanism of Dps interaction with DNA still remains enigmatic. The crucial role of lysine residues in the unstructured N-terminal tails led to the conventional point of view that Dps binds DNA without sequence or structural specificity. However, deletion of dps changed the profile of proteins in starved cells, SELEX screen revealed genomic regions preferentially bound in vitro and certain affinity of Dps for artificial branched molecules was detected by atomic force microscopy. Here we report a non-random distribution of Dps binding sites across the bacterial chromosome in exponentially growing cells and show their enrichment with inverted repeats prone to form secondary structures. We found that the Dps-bound regions overlap with sites occupied by other nucleoid proteins, and contain overrepresented motifs typical for their consensus sequences. Of the two types of genomic domains with extensive protein occupancy, which can be highly expressed or transcriptionally silent only those that are enriched with RNA polymerase molecules were preferentially occupied by Dps. In the dps-null mutant we, therefore, observed a differentially altered expression of several targeted genes and found suppressed transcription from the dps promoter. In most cases this can be explained by the relieved interference with Dps for nucleoid proteins exploiting sequence-specific modes of DNA binding. Thus, protecting bacterial cells from different stresses during exponential growth, Dps can modulate transcriptional integrity of the bacterial chromosome hampering RNA biosynthesis from some genes via competition with RNA polymerase or, vice versa, competing with inhibitors to activate transcription.
Abstract:	This work was funded by: 1. Russian Scientific Foundation (grant 14-14-00985, http://rscf.ru/node/8, PI: ONO, team members: SSA, MNT, USS (sample preparation, MiSeq sequencing, data analysis and evaluation)); 2. Russian Foundation for basic research (grant 16-34-01044, http://www.rfbr.ru/rffi/eng, PI: USS, reporter assays); 3. HHMI International Early Career Scientist Program (55007424, FAK) (http://www.hhmi.org/, FAK, HiSeq Illumina sequencing); 4. European Research Council grant agreement (335980_EinME, https://erc.europa.eu/, FAK, HiSeq Illumina sequencing); 5. Russian Foundation for Basic Research 16-04-01570 (http://www.rfbr.ru/rffi/eng) (PI ONO).
Subject(s):	-Escherichia coli -DNA-binding proteins -Bacterial genomics -DNA-directed RNA polymerases -Inverted repeat sequences
Rights:	© 2017 Antipov et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. http://creativecommons.org/licenses/by/4.0/
Document type:	Article Article - Published version
Published by:	Public Library of Science (PLoS)
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