Basic and Acidic Regions Flanking the HMG Domain of Maize HMGa Modulate the Interactions with DNA and the Self-Association of the Protein

Ritt, Christoph; Grimm, Rudi; Fernández, Silvia; Alonso, Juan C.; Grasser, Klaus D.

doi:10.1021/bi972620r

Cited by 53 publications

(95 citation statements)

References 73 publications

(142 reference statements)

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“…Purification of Proteins-Full-length and truncated recombinant maize HMGB proteins were expressed in Escherichia coli and purified by three-step column chromatography as described previously (16,17). Native HMGB proteins were isolated from maize BMS cells by 2% trichloroacetic acid extraction and subsequently purified by Resource Q chromatography as described previously (16).…”

Section: Methodsmentioning

confidence: 99%

“…Protein binding to the DNA was examined by electrophoresis on 5% polyacrylamide gels in 1ϫ TBE buffer and scanning of the gels using a Typhoon 8600 phosphorimaging device (Amersham Biosciences, Inc.) at excitation 532 nm and emission 526 nm. DNA binding analyses using a mixture of 32 P-labeled linear and circularized 78-bp DNA fragments and various concentrations of HMGB proteins by electrophoretic mobility shift assays were performed as described previously (16,17).…”

Section: Methodsmentioning

confidence: 99%

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Protein Kinase CK2 Differentially Phosphorylates Maize Chromosomal High Mobility Group B (HMGB) Proteins Modulating Their Stability and DNA Interactions

Stemmer

Schwander

Bauw

et al. 2002

Journal of Biological Chemistry

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The high mobility group (HMG) proteins of the HMGB family are architectural factors in eukaryotic chromatin, which are involved in the regulation of various DNAdependent processes. We have examined the post-translational modifications of five HMGB proteins from maize suspension cultured cells, revealing that HMGB1 and HMGB2/3, but not HMGB4 and HMGB5, are phosphorylated by protein kinase CK2. The phosphorylation sites have been mapped to the acidic C-terminal domains by analysis of tryptic peptides derived from HMGB1 and HMGB2/3 using nanospray ion trap mass spectrometry. In native HMGB1, Ser 149 is constitutively phosphorylated, whereas Ser 133 and Ser 136 are differentially phosphorylated. The functional significance of the CK2-mediated phosphorylation of HMGB proteins was analyzed by circular dichroism measurements showing that the phosphorylation increases the thermal stability of the HMGB proteins. Electrophoretic mobility shift assays demonstrate that the phosphorylation reduces the affinity of the HMGB proteins for linear DNA. The specific recognition of DNA minicircles is not affected by the phosphorylation, but a different pattern of protein-DNA complexes is formed. Collectively, these findings show that phosphorylation of residues within the acidic C-terminal domain of the HMGB proteins can modulate protein stability and the DNA binding properties of the HMGB proteins. High mobility group (HMG)1 proteins represent a heterogeneous class of small and relatively abundant chromatin-associated proteins of eukaryotes (1, 2). Proteins belonging to the subgroup of the HMGB proteins 2 (previously termed HMG1/2 proteins (3)) have in common a distinctive DNA-binding motif, termed the HMG-box domain, in which the global fold is well conserved and consists essentially of three ␣-helices arranged in an L-shape (1, 4). The HMG-box domain mediates both non-sequence-specific binding of these proteins to the minor groove of linear DNA and high affinity interactions with distorted DNA structures such as four-way junctions, minicircles, and cis-platinated DNA (2, 4, 5). In complexes with B DNA, a hydrophobic wedge on the concave surface of the HMG-box domain is inserted into the minor groove of the DNA, which contributes to the extent of DNA bending induced by the protein (5). The DNA interactions of the HMG-box domains, which occur in different plant, vertebrate, insect, and yeast HMGB proteins, are modulated by basic and acidic domains flanking the DNA-binding motif (4). HMGB proteins act as architectural components in chromatin facilitating the assembly of nucleoprotein complexes, which are involved, for instance, in the regulation of transcription and recombination (2, 4).In contrast to other eukaryotes, which usually have two or three different HMGB proteins, (higher) plants contain several HMGB proteins (Ն5 family members). The plant HMGB proteins have a single HMG-box domain, which is flanked by a basic N-terminal domain and an acidic C-terminal domain (6). Although the amino acid sequences of the HMG-box domains o...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Protein Kinase CK2 Differentially Phosphorylates Maize Chromosomal High Mobility Group B (HMGB) Proteins Modulating Their Stability and DNA Interactions

Stemmer

Schwander

Bauw

et al. 2002

Journal of Biological Chemistry

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100

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“…The binding of the HMG proteins to DNA-cellulose was measured as described previously (Ritt et al, 1998).…”

Section: Expression Purification and Characterisation Of The Proteinsmentioning

confidence: 99%

“…Chemical cross-linking of the HMG proteins was performed using the homobifunctional reagent suberic acid bis (N-hydroxysuccinimide ester) as described previously (Ritt et al, 1998).…”

Section: Expression Purification and Characterisation Of The Proteinsmentioning

confidence: 99%

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Four differently chromatin‐associated maize HMG domain proteins modulate DNA structure and act as architectural elements in nucleoprotein complexes

et al. 1998

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SummaryIn contrast to other eukaryotes which usually express two closely related HMG1-like proteins, plant cells have multiple relatively variable proteins of this type. A systematic analysis of the DNA-binding properties of four chromosomal HMG domain proteins from maize revealed that they bind linear DNA with similar affinity. HMGa, HMGc1/2 and HMGd specifically recognise diverse DNA structures such as DNA mini-circles and supercoiled DNA. They induce DNA-bending, and constrain negative superhelical turns in DNA. In the presence of DNA, the HMG domain proteins can self-associate, whereas they are monomeric in solution. The maize HMG1-like proteins have the ability to facilitate the formation of nucleoprotein structures to different extents, since they can efficiently replace a bacterial chromatin-associated protein required for the site-specific β-mediated recombination. A variable function of the HMG1-like proteins is indicated by their differential association with maize chromatin, as judged by their 'extractability' from chromatin with spermine and ethidium bromide. Collectively, these findings suggest that the various plant chromosomal HMG domain proteins could be adapted to act in different nucleoprotein structures in vivo.

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The self‐association of HMGB1 and its possible role in the binding to DNA and cell membrane receptors

et al. 2017

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Edited by Miguel De la RosaHigh mobility group box 1 (HMGB1), a chromatin protein, interacts with DNA and controls gene expression. However, when HMGB1 is released from apoptotic or damaged cells, it triggers proinflammatory reactions by interacting with various receptors, mainly receptor for advanced glycation end-products (RAGE) and toll-like receptors (TLRs). The self-association of HMGB1 has been found to be crucial for its DNA-related biological functions. It is influenced by several factors, such as ionic strength, pH, specific divalent metal cations, redox environment and acetylation. This self-association may also play a role in the interaction with RAGE and TLRs and the concomitant inflammatory responses. Future studies should address the potential role of HMGB1 self-association on its interactions with DNA, RAGE and TLRs, as well as the influence of physicochemical factors in different cellular environments on these interactions.

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Basic and Acidic Regions Flanking the HMG Domain of Maize HMGa Modulate the Interactions with DNA and the Self-Association of the Protein

Cited by 53 publications

References 73 publications

Protein Kinase CK2 Differentially Phosphorylates Maize Chromosomal High Mobility Group B (HMGB) Proteins Modulating Their Stability and DNA Interactions

Protein Kinase CK2 Differentially Phosphorylates Maize Chromosomal High Mobility Group B (HMGB) Proteins Modulating Their Stability and DNA Interactions

Four differently chromatin‐associated maize HMG domain proteins modulate DNA structure and act as architectural elements in nucleoprotein complexes

The self‐association of HMGB1 and its possible role in the binding to DNA and cell membrane receptors

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