PMID:19477901

Citation	Werlang, IC, Schneider, CZ, Mendonça, JD, Palma, MS, Basso, LA and Santos, DS (2009) Identification of Rv3852 as a nucleoid-associated protein in Mycobacterium tuberculosis. Microbiology (Reading, Engl.) 155:2652-63
Abstract	Tuberculosis remains the major cause of mortality due to a bacterial pathogen, Mycobacterium tuberculosis. The molecular mechanisms of infection and persistence have not been completely elucidated for this pathogen. Studies involving nucleoid-associated proteins (NAPs), which have been related to the control and influence of virulence genes in pathogenic bacteria, can help unveil the virulence process of M. tuberculosis. Here, we describe the initial characterization of an ORF for an M. tuberculosis putative NAP. The Rv3852 gene was cloned and expressed, and its product purified to homogeneity. A qualitative protein-DNA binding assay was carried out by gel-retardation and the protein affinity for specific DNA sequences was assessed quantitatively by surface plasmon resonance (SPR). A stoichiometry of 10 molecules of monomeric protein per molecule of DNA was determined. The monophasic apparent dissociation rate constant values increased to a saturable level as a function of protein concentration, yielding two limiting values for the molecular recognition of proU2 DNA. A protein-DNA binding mechanism is proposed. In addition, functional complementation studies with an Escherichia coli hns mutant reinforce the likelihood that the Rv3852 protein represents a novel NAP in M. tuberculosis.
Links	PubMed Online version:10.1099/mic.0.030148-0
Keywords	Amino Acid Sequence; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Cloning, Molecular; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Electrophoresis, Agar Gel; Electrophoretic Mobility Shift Assay; Escherichia coli/metabolism; Molecular Sequence Data; Mycobacterium tuberculosis/genetics; Mycobacterium tuberculosis/metabolism; Open Reading Frames; Recombinant Proteins/biosynthesis; Recombinant Proteins/genetics; Sequence Alignment; Sequence Analysis, Protein
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
MYCTU:P96225		GO:0042803: protein homodimerization activity	IPI: Inferred from Physical Interaction: UniProtKB:P96225	F	Seeded From UniProt	complete
MYCTU:P96225		GO:0006355: regulation of transcription, DNA-templated	IDA: Inferred from Direct Assay:	P	Seeded From UniProt	complete
MYCTU:P96225		GO:0003677: DNA binding	IDA: Inferred from Direct Assay:	F	Seeded From UniProt	complete

References

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