Purification and functional analysis of recombinant Acholeplasma laidlawii histone-like HU protein

Levitskiy, S. A.; Sycheva, A.M.; Kharlampieva, Daria; Oberto, Jacques; Kamashev, Dmitri; Serebryakova, Marina V.; Moshkovskii, Sergei A.; Лазарев, В. Н.; Govorun, Vadim M.

doi:10.1016/j.biochi.2011.03.005

Cited by 12 publications

(10 citation statements)

References 39 publications

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“…We have identified about a third of all possible intracellular metabolites predicted by genome annotation and compared the metabolite content of three Mollicute species. Metabolomic data analysis is based on a previous proteogenomic study performed in our lab for all three bacterial species [3], [10], [11], [12]. The high-throughput-omics analysis, including transcriptomics, proteomics, and complemented with the metabolomic study performed in this work, will improve our comprehensive understanding how the three closely-related Mollicute species are regulated under different conditions [13].…”

Section: Introductionmentioning

confidence: 99%

Metabolomic Analysis of Three Mollicute Species

et al. 2014

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We present a systematic study of three bacterial species that belong to the class Mollicutes, the smallest and simplest bacteria, Spiroplasma melliferum, Mycoplasma gallisepticum, and Acholeplasma laidlawii. To understand the difference in the basic principles of metabolism regulation and adaptation to environmental conditions in the three species, we analyzed the metabolome of these bacteria. Metabolic pathways were reconstructed using the proteogenomic annotation data provided by our lab. The results of metabolome, proteome and genome profiling suggest a fundamental difference in the adaptation of the three closely related Mollicute species to stress conditions. As the transaldolase is not annotated in Mollicutes, we propose variants of the pentose phosphate pathway catalyzed by annotated enzymes for three species. For metabolite detection we employed high performance liquid chromatography coupled with mass spectrometry. We used liquid chromatography method - hydrophilic interaction chromatography with silica column - as it effectively separates highly polar cellular metabolites prior to their detection by mass spectrometer.

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Section: Introductionmentioning

confidence: 99%

Metabolomic Analysis of Three Mollicute Species

et al. 2014

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“…This led to the conclusion that mutation rate is elevated in mycoplasmas [14]; however, it was later shown that it does not differ from the one in E. coli [15,16]. It was shown recently that histone-like protein HU (Hup2) in M. gallisepticum is able to recognize mispaired bases in DNA in contrast to HU from Acholeplasma laidlawii with functional MMR [17,18]. It can also compensate for inactivation of hupAB proteins in E. coli [17].…”

Section: Introductionmentioning

confidence: 99%

DNA repair in Mycoplasma gallisepticum

et al. 2013

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BackgroundDNA repair is essential for the maintenance of genome stability in all living beings. Genome size as well as the repertoire and abundance of DNA repair components may vary among prokaryotic species. The bacteria of the Mollicutes class feature a small genome size, absence of a cell wall, and a parasitic lifestyle. A small number of genes make Mollicutes a good model for a “minimal cell” concept.ResultsIn this work we studied the DNA repair system of Mycoplasma gallisepticum on genomic, transcriptional, and proteomic levels. We detected 18 out of 22 members of the DNA repair system on a protein level. We found that abundance of the respective mRNAs is less than one per cell. We studied transcriptional response of DNA repair genes of M. gallisepticum at stress conditions including heat, osmotic, peroxide stresses, tetracycline and ciprofloxacin treatment, stationary phase and heat stress in stationary phase.ConclusionsBased on comparative genomic study, we determined that the DNA repair system M. gallisepticum includes a sufficient set of proteins to provide a cell with functional nucleotide and base excision repair and mismatch repair. We identified SOS-response in M. gallisepticum on ciprofloxacin, which is a known SOS-inducer, tetracycline and heat stress in the absence of established regulators. Heat stress was found to be the strongest SOS-inducer. We found that upon transition to stationary phase of culture growth transcription of DNA repair genes decreases dramatically. Heat stress does not induce SOS-response in a stationary phase.

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“…In bacteria, the nucleoid-associated protein (NAP) HU is one of the most abundant basic proteins [1][2][3][4]. The other major NAPs include factor for inversion stimulation, heat-stable nucleoid-structural protein, integration host factor, and a stationary-phase specific DNA-binding protein, Dps [5][6][7][8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

“…The other major NAPs include factor for inversion stimulation, heat-stable nucleoid-structural protein, integration host factor, and a stationary-phase specific DNA-binding protein, Dps [5][6][7][8][9][10][11]. HU plays a dual role in helping to maintain nucleoid architecture and as a regulator of various processes related to DNA metabolism [1][2][3][4]. HU is highly conserved in its heterodimeric form (HUab) or in its homodimeric form (HUaa or HUbb) [6][7][8][9][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Synergy between the N‐terminal and C‐terminal domains of Mycobacterium tuberculosis HupB is essential for high‐affinity binding, DNA supercoiling and inhibition of RecA‐promoted strand exchange

Sharadamma¹,

Khan²,

Kumar³

et al. 2011

The FEBS Journal

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The occurrence of DNA architectural proteins containing two functional domains derived from two different architectural proteins is an interesting emerging research theme in the field of nucleoid structure and function. Mycobacterium tuberculosis HupB, unlike Escherichia coli HU, is a two-domain protein that, in the N-terminal region, shows broad sequence homology with bacterial HU. The long C-terminal extension, on the other hand, contains seven PAKK/KAAK motifs, which are characteristic of the histone H1/H5 family of proteins. In this article, we describe several aspects of HupB function, in comparison with its truncated derivatives lacking either the C-terminus or N-terminus. We found that HupB binds a variety of DNA repair and replication intermediates with K(d) values in the nanomolar range. By contrast, the N-terminal fragment of M. tuberculosis HupB (HupB(MtbN)) showed diminished DNA-binding activity, with K(d) values in the micromolar range, and the C-terminal domain was completely devoid of DNA-binding activity. Unlike HupB(MtbN) , HupB was able to constrain DNA in negative supercoils and introduce negative superhelical turns into relaxed DNA. Similarly, HupB exerted a robust inhibitory effect on DNA strand exchange promoted by cognate and noncognate RecA proteins, whereas HupB(MtbN), even at a 50-fold molar excess, had no inhibitory effect. Considered together, these results suggest that synergy between the N-terminal and C-terminal domains of HupB is essential for its DNA-binding ability, and to modulate the topological features of DNA, which has implications for processes such as DNA compaction, gene regulation, homologous recombination, and DNA repair.

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Purification and functional analysis of recombinant Acholeplasma laidlawii histone-like HU protein

Cited by 12 publications

References 39 publications

Metabolomic Analysis of Three Mollicute Species

Metabolomic Analysis of Three Mollicute Species

DNA repair in Mycoplasma gallisepticum

Synergy between the N‐terminal and C‐terminal domains of Mycobacterium tuberculosis HupB is essential for high‐affinity binding, DNA supercoiling and inhibition of RecA‐promoted strand exchange

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