A Role of Basic Residues and the Putative Intercalating Phenylalanine of the HMG-1 Box B in DNA Supercoiling and Binding to Four-way DNA Junctions

Štros, Michal; Muselı́ková, Eva

doi:10.1074/jbc.m007167200

Cited by 46 publications

(52 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These residues were targeted for mutagenesis based on sequence and structural analysis indicating that they comprise part of a "hydrophobic wedge" in their respective HMG-box domains that mediates contact with the minor groove of DNA (10). Phenylalanine 103, for example, is thought to intercalate between DNA base pairs, promoting DNA bending; mutation of this residue impairs the DNA supercoiling activity of box B (25). Mutagenesis also replaces a highly conserved arginine residue present in both HMG-boxes (Arg 24 in box A, and Arg 110 in box B); substitution of this residue in either HMG-box reduces the DNA binding activity of the domain (25,26).…”

Section: Resultsmentioning

confidence: 99%

“…Phenylalanine 103, for example, is thought to intercalate between DNA base pairs, promoting DNA bending; mutation of this residue impairs the DNA supercoiling activity of box B (25). Mutagenesis also replaces a highly conserved arginine residue present in both HMG-boxes (Arg 24 in box A, and Arg 110 in box B); substitution of this residue in either HMG-box reduces the DNA binding activity of the domain (25,26). To test whether the orientation of the two HMG-box domains relative to one another is an important determinant for the ability of HMGB1 to stimulate RAG complex binding and cleavage activity, we also prepared a version of HMGB1 in which the positions of the two HMG-box domains are inverted or "shuffled."…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Both High Mobility Group (HMG)-boxes and the Acidic Tail of HMGB1 Regulate Recombination-activating Gene (RAG)-mediated Recombination Signal Synapsis and Cleavage in Vitro

Bergeron

Madathiparambil

Swanson

2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

RAG-1 and RAG-2 initiate V(D)J recombination through synapsis and cleavage of a 12/23 pair of V(D)J recombination signal sequences (RSS). RAG-RSS complex assembly and activity in vitro is promoted by high mobility group proteins of the "HMG-box" family, exemplified by HMGB1. How HMGB1 stimulates the DNA binding and cleavage activity of the RAG complex remains unclear. HMGB1 contains two homologous HMGbox DNA binding domains, termed A and B, linked by a stretch of basic residues to a highly acidic C-terminal tail. To identify determinants of HMGB1 required for stimulation of RAG-mediated RSS binding and cleavage, we prepared an extensive panel of mutant HMGB1 proteins and tested their ability to augment RAG-mediated RSS binding and cleavage activity. Using a combination of mobility shift and in-gel cleavage assays, we find that HMGB1 promotes RAG-mediated cleavage largely through the activity of box B, but optimal stimulation requires a functional A box tethered in the correct orientation. Box A or B mutants fail to promote RAG synaptic complex formation, but this defect is alleviated when the acidic tail is removed from these mutants.During lymphocyte development, antigen receptor genes undergo a series of DNA rearrangements to generate functional exons encoding the antigen binding domains of these receptors (1). This rearrangement process, termed V(D)J recombination, is initiated when two lymphoid cell-specific proteins, called recombination-activating gene (RAG) 1 -1 and RAG-2, bring two gene segments into close proximity and then introduce a DNA double-strand break (DSB) at the end of each coding segment.Adjacent to each coding segment lies a recombination signal sequence (RSS) that serves as the binding site for the RAG-1/2 protein complex (hereafter termed the "RAG complex") and directs the location of cleavage. The RSS contains a conserved heptamer and nonamer motif separated by either 12 or 23 bp of relatively nonconserved sequence (12-RSS and 23-RSS, respectively). Productive exon assembly is promoted by the 12/23 rule, a restriction that limits rearrangement to RSSs whose spacing between the heptamer and nonamer is different. RAGmediated cleavage of RSS pairs produces four DNA ends: two blunt, 5Ј-phosphorylated signal ends and two coding ends terminating in DNA hairpin structures (2-4). These reaction products originate from a two-step cleavage reaction in which the RSS is first nicked at its 5Ј-end, and then the resulting 3Ј-OH is covalently linked to the bottom strand by direct transesterification (5, 6). After DNA cleavage, signal ends are generally ligated together to form precise signal joints, but coding ends, being sealed as DNA hairpins, are first resolved and then processed and joined to create coding joints in which nucleotides are frequently gained or lost at the junction. DNA hairpin opening is most likely catalyzed by a complex containing Artemis and the catalytic subunit of the DNA-dependent protein kinase (DNA-PKcs) (7). Signal and coding joint formation is mediated by ubiquitously expr...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Both High Mobility Group (HMG)-boxes and the Acidic Tail of HMGB1 Regulate Recombination-activating Gene (RAG)-mediated Recombination Signal Synapsis and Cleavage in Vitro

Bergeron

Madathiparambil

Swanson

2005

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…2B). Four of the six residues, Lys-87, Lys-90, Lys-96, and Arg-97, also contribute to DNA binding (30,31). However, DNA binding requires at least four additional charged residues in the A-box and six in the B-box (32).…”

Section: Discussionmentioning

confidence: 99%

Heparan Sulfate Is Essential for High Mobility Group Protein 1 (HMGB1) Signaling by the Receptor for Advanced Glycation End Products (RAGE)

Young

Song

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background:The cytokine function of the danger-associated molecular pattern protein HMGB1 is mediated through RAGE. Results: HMGB1-RAGE signaling depends on heparan sulfate, but heparan sulfate binding to HMGB1 is dispensable. Conclusion: Heparan sulfate is essential for HMGB1 signaling because RAGE binds heparan sulfate. Significance: Perturbing heparan sulfate may be a novel strategy to alter RAGE-dependent signaling.

show abstract

“…Isolation of HMGB1-HMGB1 was isolated under nondenaturing conditions from calf thymus and highly purified to near homogeneity on fast protein liquid chromatography as described previously (6,24).…”

Section: Methodsmentioning

confidence: 99%

HMGB1 and HMGB2 Cell-specifically Down-regulate the p53- and p73-dependent Sequence-specific Transactivation from the Human Bax Gene Promoter

Štros¹,

Ozaki²,

Bačı́ková³

et al. 2002

Journal of Biological Chemistry

Self Cite

170

133

View full text Add to dashboard Cite

A Role of Basic Residues and the Putative Intercalating Phenylalanine of the HMG-1 Box B in DNA Supercoiling and Binding to Four-way DNA Junctions

Cited by 46 publications

References 53 publications

Both High Mobility Group (HMG)-boxes and the Acidic Tail of HMGB1 Regulate Recombination-activating Gene (RAG)-mediated Recombination Signal Synapsis and Cleavage in Vitro

Both High Mobility Group (HMG)-boxes and the Acidic Tail of HMGB1 Regulate Recombination-activating Gene (RAG)-mediated Recombination Signal Synapsis and Cleavage in Vitro

Heparan Sulfate Is Essential for High Mobility Group Protein 1 (HMGB1) Signaling by the Receptor for Advanced Glycation End Products (RAGE)

HMGB1 and HMGB2 Cell-specifically Down-regulate the p53- and p73-dependent Sequence-specific Transactivation from the Human Bax Gene Promoter

Contact Info

Product

Resources

About