| 1993 |
HMGB2 (HMG2) bends DNA extremely efficiently, forming circles as small as 66 bp, and can substitute for prokaryotic HU protein in promoting assembly of complex nucleoprotein invertasome structures, with HMG box domain B being sufficient for both bending and invertasome assembly. |
Ligase-mediated circularization assay, invertasome assembly assay, partial proteolytic digestion domain analysis |
Genes & development |
High |
8339930
|
| 1978 |
HMG2 reduces the linking number of circular DNA when covalent closure occurs in its presence, indicating the protein unwinds the DNA double helix or induces supercoiling. |
Topological winding number assay with circular DNA and DNA ligase |
Science |
High |
628842
|
| 1979 |
HMG2 unwinds the DNA double helix by local denaturation of base pairs; HMG2 shows higher affinity for single-stranded than double-stranded DNA in the presence of salt; the net unwinding angle is 26° per molecule of HMG2. |
Melting absorption technique, competition unwinding experiments measuring topological winding number |
Nucleic acids research |
High |
226939
|
| 1995 |
HMG2 activates transcription in a defined in vitro RNA polymerase II system by stabilizing an activated conformation of the TFIID-TFIIA-promoter complex; activation requires TFIIA and TAF-containing TFIID, and results in slower TFIIB dissociation from the preinitiation complex. |
Defined in vitro transcription reconstitution assay with purified factors, order-of-addition experiments, titration experiments |
Genes & development |
High |
7797075
|
| 1994 |
HMG2 acts as a basal repressor of class II gene transcription by inhibiting steps after TBP-TFIIA-promoter complex assembly but before the fourth phosphodiester bond is formed; a TFIIH-associated factor counteracts this repression in an ATP/dATP-dependent manner. |
In vitro transcription assay with purified factors, order-of-addition experiments, antibody depletion of TFIIH, ATP analog inhibition |
Molecular and cellular biology |
High |
8007973
|
| 1999 |
The RAG1 homeodomain directly interacts with both HMG boxes of HMG2; this interaction facilitates RAG1/2 binding to recombination signal sequences (RSS) by promoting high-affinity binding to the nonamer motif, and HMG2 significantly enhances bending of the 23RSS. |
Protein-protein interaction assay, circular permutation DNA bending assay, electrophoretic mobility shift assay, in vivo V(D)J recombination assay in transfected cells |
Molecular and cellular biology |
High |
10490593
|
| 1999 |
HMG2 is stably incorporated into the RAG1/RSS complex; it can increase the affinity of RAG1 for the RSS even in the absence of RAG2. Core RAG1 exists as a dimer both free in solution and as the minimal species bound to the RSS. |
Electrophoretic mobility shift assay (EMSA), recombinant protein expression, zinc analysis, cleavage activity assay |
Nucleic acids research |
High |
10390537
|
| 2002 |
HMG2 is a component of the SET complex (270–420 kDa endoplasmic reticulum-associated complex containing SET, pp32, and APE); it coprecipitates with SET and colocalizes with SET at the endoplasmic reticulum in the cytoplasm. Cytoplasmic HMG2 mediates DNA binding and bending activity within the SET complex. HMG2 is a physiologically relevant granzyme A substrate; granzyme A cleaves HMG2 after Lys65 within HMG box A, destroying its DNA binding and bending functions. |
Co-immunoprecipitation, confocal microscopy colocalization, in vitro granzyme A cleavage assay, DNA binding/bending assays, single-stranded DNA nicking assay in isolated nuclei |
Molecular and cellular biology |
High |
11909973
|
| 2001 |
HMGB2 physically interacts with p73α and p73β (pull-down assay); both HMG box domains A and B of HMGB1 interact with p73α; endogenous or ectopically expressed HMGB2 cell-specifically inhibits p73α/β- and p53-dependent transactivation from the Bax gene promoter in SAOS-2 cells, while stimulating it in H1299 cells. |
Pull-down assay, gel-shift assay (EMSA), transient transfection with luciferase reporter, antisense knockdown |
The Journal of biological chemistry |
High |
11748232
|
| 2001 |
Hmgb2 knockout mice are viable but male Hmgb2-/- mice have reduced fertility associated with Sertoli and germ cell degeneration and immotile spermatozoa; in adult mice Hmgb2 is restricted to lymphoid organs and testes, though widely expressed during embryogenesis, indicating a specialized role in germ cell differentiation. |
Gene knockout (Hmgb2-/- mice), histological analysis, immunohistochemistry, fertility assay |
Development |
High |
11262228
|
| 2003 |
HMGB1 and HMGB2 are present in mitotic cells in two forms—free and chromatin-associated—that rapidly exchange; two sites encompassing HMG-box A and B are responsible for mitotic chromosome binding; previous immunofluorescence reports of dissociation from chromosomes during mitosis were artifacts of cell permeabilization or chemical fixation. |
Live-cell imaging with GFP/DsRed-tagged proteins, fluorescence recovery after photobleaching (FRAP), permeabilization controls |
Molecular biology of the cell |
High |
12925773
|
| 1997 |
Nuclear accumulation of HMG2 is mediated by basic regions interspaced with the HMG box DNA-binding sequence (a novel nuclear localization signal distinct from classic basic clusters); retention within the nucleus requires the acidic carboxyl terminus, as shown by interspecies heterokaryon assay. |
HMG2-β-galactosidase fusion protein expression in COS-7 cells, deletion analysis, interspecies heterokaryon assay |
Biochemistry |
High |
9166769
|
| 1998 |
HMG2 activates the nuclease activity of DFF40 (a caspase-3-activated apoptotic DNA fragmentation factor); bacterially expressed HMG2 enhances DFF40 nuclease activity in vitro, suggesting HMG proteins augment apoptotic DNA fragmentation through chromosomal structural changes. |
In vitro nuclease activity assay with purified recombinant proteins |
Biochemical and biophysical research communications |
Medium |
9784391
|
| 1999 |
HMG box B of HMG2 is the primary domain for DNA recognition and conformational changes (supercoiled DNA binding and DNA unwinding); box A requires the flanking basic linker region or box B to achieve full activity; Phe-102 in box B is predicted to intercalate into the base stack, whereas Ala-16 in box A is too small to intercalate. |
Gel retardation assay, DNA supercoiling assay, surface plasmon resonance (SPR), deletion/domain expression in E. coli, computer modeling |
Biochemistry |
High |
9888798
|
| 2005 |
Isolated HMG box A from HMGB2 induces an average DNA bend angle of ~114° (at 50 mM Na+) via a flexible hinge mechanism at low concentrations; at higher concentrations, a cooperative filament binding mode is observed, distinct from the hinge mode. |
Single-molecule optical tweezers force-extension measurement of DNA |
Biophysical journal |
High |
15833996
|
| 2009 |
HMGB1 and HMGB2 upregulate cellular expression of topoisomerase IIα; HMGB1/2 knockdown significantly decreases topo IIα mRNA and protein levels; the mechanism involves HMGB1/2 modulating binding of transcription factor NF-Y to the topo IIα promoter, and this effect is dependent on pRb status (active in pRb-negative cells). |
siRNA knockdown, luciferase reporter assay (topo IIα promoter), western blot, RT-PCR, pRb overexpression rescue experiment |
Nucleic acids research |
Medium |
19223331
|
| 2009 |
Age-related loss of HMGB2 in articular cartilage superficial zone is associated with reduced cellularity due to increased chondrocyte apoptosis; Hmgb2-/- mice show earlier onset and more severe osteoarthritis with increased cell death preceding glycosaminoglycan depletion. |
Hmgb2 knockout mice (Hmgb2-/-), histological analysis of cartilage, in vitro apoptosis assay of Hmgb2-/- chondrocytes |
PNAS |
High |
19139395
|
| 2009 |
HMGB2 enhances binding of Lef-1 to its target sequence and potentiates transcriptional activation of the Lef-1-β-catenin complex; the HMG domain within HMGB2 is required for interaction with Lef-1; HMGB2 and Wnt/β-catenin pathway co-localize in the superficial zone, and conditional deletion of β-catenin in chondrocytes induces apoptosis. |
Co-immunoprecipitation, ChIP, transcription reporter assay, conditional knockout of β-catenin in chondrocytes, immunofluorescence |
PNAS |
High |
19805379
|
| 2009 |
Acetylated forms of HMGB1 and HMGB2 (in vivo acetylated) acquire the ability to bind to nucleosome core particles (not shown for non-acetylated forms); acetylation increases binding to linker DNA-containing nucleosomes and enhances co-remodeling activity with SWI/SNF and RSC; acetylated HMGB1/2 also enhance SWI/SNF binding to nucleosomes without affecting ATPase activity. |
Nucleosome binding assay, nucleosome mobilization/sliding assay with SWI/SNF and RSC, ATPase assay |
Biochemistry |
Medium |
19522541
|
| 2009 |
HMGB2 directly binds to the GFI1B promoter in vivo and up-regulates its transactivation, most likely by enhancing the binding of Oct-1 and GATA-1 to the GFI1B promoter; knockdown of HMGB2 in hematopoietic progenitor cells decreases Gfi-1B expression and impairs erythroid differentiation. |
ChIP (chromatin immunoprecipitation), luciferase reporter assay, siRNA knockdown, erythroid differentiation assay |
Blood |
High |
19965638
|
| 2011 |
HMGB2 suppresses chondrogenic differentiation: lentiviral HMGB2 transduction of MSCs inhibits Col2a1 and Col10a1 expression, while Hmgb2-/- MSCs show enhanced Col10a1 and Runx2 expression; HMGB2 negatively regulates the stimulatory effect of Wnt/β-catenin signaling on the Runx2 proximal promoter. |
Lentiviral transduction, Hmgb2-/- MSC analysis, RT-PCR, luciferase reporter assay (Runx2 promoter), western blot |
The Journal of biological chemistry |
High |
21890638
|
| 2012 |
HMGB2 knockdown sensitizes colorectal cancer cells to ionizing radiation by increasing DNA damage and reducing DNA damage repair efficiency; p53 transcriptionally downregulates HMGB2 expression after radiation exposure, as shown by luciferase reporter assay (p53 reduces HMGB2 promoter activity) and by Nutlin-3/Tet-On p53 induction. |
shRNA knockdown, clonogenic survival assay, comet assay (DNA damage), luciferase reporter assay (HMGB2 promoter), Nutlin-3 treatment, Tet-On p53 induction system |
Cancer biology & therapy |
High |
23255232
|
| 2013 |
Oct4 post-translational modifications (phosphorylation) promote Akt activation and interaction of Oct4 with Hmgb2 and the SET complex, which preserves H3K27me3 marks in daughter cells and maintains pluripotency gene expression in murine embryonic stem cells. |
Co-immunoprecipitation, western blot (H3K27me3 ChIP), phosphorylation mutant analysis, Akt inhibition |
Stem cells |
Medium |
23495099
|
| 2013 |
Hmgb2-/- mice exhibit subventricular zone hyperproliferation with increased numbers of neural stem cells and increased newly born neurons in the olfactory bulb; loss of HMGB2 is associated with upregulation of p21 and NCAM and downregulation of Oct4 in the SVZ. |
Hmgb2-/- mouse analysis, BrdU/Ki67 immunohistochemistry, western blot, neurosphere assays |
PloS one |
Medium |
24391977
|
| 2014 |
HMGB2 forms a complex with SRC-1 and estrogen receptor (ER) at promoter regions of target genes in tamoxifen-resistant breast cancer cells; HMGB2 ChIPseq reveals distinct binding patterns between endocrine-sensitive and -resistant contexts; the HMGB2/SRC-1/ER complex targets the RNA helicase DDX18, whose modulation directly affects growth of tamoxifen-resistant cells. |
Co-immunoprecipitation (SRC-1/HMGB2), ChIP-seq (HMGB2), gene expression analysis, DDX18 knockdown proliferation assay |
Oncogene |
High |
25284587
|
| 2015 |
HMGB2 directly binds to Lrp1-AS (a natural antisense lncRNA) and this interaction inhibits HMGB2's ability to enhance Srebp1a-dependent transcription of Lrp1; short oligonucleotides targeting Lrp1-AS inhibit the lncRNA-HMGB2 interaction and increase Lrp1 expression. |
RNA immunoprecipitation (RIP), in vitro binding assay, luciferase reporter assay (Lrp1 promoter), antisense oligonucleotide treatment |
Cell reports |
High |
25937287
|
| 2016 |
HMGB2 preferentially localizes to SASP gene loci during senescence; loss of HMGB2 allows spreading of repressive heterochromatin into SASP gene loci, causing SASP gene loci to be incorporated into SAHFs and blunting SASP gene expression; HMGB2 thus acts as a boundary factor preventing heterochromatin spreading. |
ChIP-seq (HMGB2, H3K9me2/3, H3K27me3), immunofluorescence, siRNA knockdown, gene expression analysis |
The Journal of cell biology |
High |
27799366
|
| 2016 |
HMGB2 regulates myoblast proliferation and differentiation; knockdown of HMGB2 inhibits myoblast proliferation and stimulates differentiation by reducing Myf5 and cyclin A2 protein (not mRNA) levels; HMGB2 acts through transcriptional upregulation of IGF2BP2, an RNA-binding protein that enhances Myf5 translation and stabilizes cyclin A2 mRNA. In vivo HMGB2 depletion attenuates muscle regeneration and reduces satellite cell numbers. |
siRNA knockdown, lentiviral overexpression, western blot, RT-PCR, in vivo muscle injury model, ChIP, immunostaining |
Journal of cell science |
High |
27672022
|
| 2018 |
Nuclear depletion of HMGB2 occurs early upon senescence entry and is sufficient to cause senescence-associated CTCF clustering and loop reshuffling; ectopic HMGB2 expression rescues these effects; HMGB2 loss is thus a mechanistic driver of 3D genome reorganization at senescence entry. |
Hi-C (3D genome), single-cell and population transcriptomics, live imaging, in silico modeling, HMGB2 knockdown, HMGB2 overexpression rescue across three cell types |
Molecular cell |
High |
29706538
|
| 2017 |
HMGB2 promotes neointimal hyperplasia and VSMC proliferation/migration by inducing reactive oxygen species through increased p47phox phosphorylation; these effects are mediated through RAGE (receptor for advanced glycation end products) but not TLR4, as shown by knockdown/knockout experiments. |
Hmgb2-/- mouse femoral artery injury model, perivascular HMGB2 administration, siRNA knockdown of p47phox and RAGE/TLR4, ROS measurement, VSMC proliferation/migration assay |
Arteriosclerosis, thrombosis, and vascular biology |
High |
28183701
|
| 2009 |
HMGB2 stabilizes p53 protein in HPV-positive HeLa cells by interfering with HPV E6/E6AP-mediated ubiquitination and proteasomal degradation of p53; HMGB2 overexpression in HeLa cells causes p53 accumulation, G1 cell cycle arrest, and decreased proliferation; this effect is specific to HPV-positive cells. |
siRNA knockdown, overexpression, western blot (p53 protein level), ubiquitination assay, FACS cell cycle analysis |
Cancer letters |
Medium |
20036050
|
| 2019 |
HMGB2 is required for the differentiation and maintenance of stem-like progenitor exhausted CD8+ T cells (Tpex) during chronic viral infection and in tumors through epigenetic and transcriptional programming; Hmgb2-/- CD8+ T cells cannot sustain Tpex differentiation and long-term survival during persistent antigen despite expressing TCF-1 and TOX. |
Hmgb2-/- mice, chronic LCMV infection model, tumor models, flow cytometry, ATAC-seq, RNA-seq, single-cell analysis |
Nature communications |
High |
37704621
|
| 2019 |
HMGB2 is a transcriptional suppressor of latexin; HMGB2 knockdown increases latexin expression and decreases hematopoietic stem cell (HSC) number and regeneration capacity in vivo; concomitant latexin blockade rescues HSC numbers; a functional SNP (rs31528793) in the latexin promoter differentially binds HMGB2 and affects promoter activity. |
DNA pull-down with mass spectrometry (identification), HMGB2 knockdown in mice, in vivo HSC number and regeneration assays, luciferase reporter assay (latexin promoter with SNP variants), western blot/RT-PCR |
Haematologica |
High |
31171637
|
| 2019 |
HMGB2 is a negative regulator of telomerase activity in human embryonic stem cells and neuroectodermal cells; HMGB2 knockdown stimulates telomerase activity potentially through activation of PI3K/AKT/GSK3β/β-catenin pathways and augmented TERT transcription, while HMGB1 has the opposite (enhancing) effect on telomerase. |
Inducible shRNA knockdown in hESCs and neuroectodermal cells, telomerase activity assay (TRAP), RT-PCR for TERT/TERC, signaling pathway inhibition |
FASEB journal |
Medium |
31661640
|
| 2021 |
HMGB2 mediates adipogenesis by binding to the C/EBPβ promoter at the sequence 'GGGTCTCAC' specifically during mitotic clonal expansion (MCE) stage, enhancing C/EBPβ expression; exogenous C/EBPβ rescues adipogenic impairment caused by HMGB2 depletion; Hmgb2-/- mice have decreased adipose tissue mass. |
Hmgb2-/- mice, ChIP (HMGB2 binding to C/EBPβ promoter), lentiviral overexpression/knockdown, C/EBPβ rescue experiment, qPCR, western blot, in vivo adipose tissue analysis |
Cell death & disease |
High |
34215724
|
| 2024 |
NAT10-mediated N4-acetylcytidine (ac4C) modification within the coding sequence (CDS) of HMGB2 mRNA enhances HMGB2 translation by facilitating binding of eukaryotic elongation factor 2 (eEF2) to the ac4C sites; this NAT10-ac4C/eEF2-HMGB2 axis promotes HCC proliferation and metastasis. |
acRIP-seq, RNA-seq, ribosome profiling, RNA immunoprecipitation, RNA pull-down, mass spectrometry, site-specific mutation, CETSA/DARTS drug binding assay, in vitro and in vivo HCC models |
Cancer communications |
High |
39030964
|
| 2024 |
HMGB2 secretion from the cell nucleus into the extracellular milieu is required for oxaliplatin-induced calreticulin (CRT) translocation to the plasma membrane (a required step for immunogenic cell death); nuclear export of HMGB2 is controlled by XPO1 (CRM1); XPO1 inhibition causes nuclear HMGB2 accumulation and inhibits CRT translocation and ferroptosis; exogenous cell-targeted HMGB2 (CT-HMGB2) is three orders of magnitude more potent than oxaliplatin at inducing CRT translocation. |
XPO1 inhibition (selinexor), nuclear fractionation, confocal microscopy (CRT translocation), ferroptosis assay, exogenous CT-HMGB2 protein treatment, flow cytometry |
Communications biology |
High |
39354146
|
| 2022 |
Spatiotemporal expression of HMGB2 regulates hepatocyte proliferation and cell size during liver regeneration; Hmgb2-/- mice show significantly delayed hepatocyte proliferation with decreased cyclin D1 and cyclin B1 mRNA, and larger hepatocytes after partial hepatectomy, indicating hepatocyte hypertrophy compensates for reduced proliferation. |
Hmgb2-/- mice, 70% partial hepatectomy model, immunohistochemistry (Ki67, PCNA), flow cytometry, qPCR (cyclin D1/B1), siRNA knockdown in vitro |
Scientific reports |
High |
35831365
|
| 2018 |
Loss of HMGB2 in cardiomyocytes causes AKT inactivation and decreased SERCA2a activity; Hmgb2-/- mice show baseline cardiac dysfunction and worsened cardiac dysfunction after TAC (pressure overload), demonstrating HMGB2 has a cardioprotective role by maintaining AKT signaling and calcium pump activity. |
Hmgb2-/- mice, TAC surgery, echocardiography, western blot (AKT phosphorylation, SERCA2a), cardiac function measurements |
Circulation journal |
Medium |
30487376
|
| 2018 |
HMGB2 overexpression promotes ischemia/reperfusion-induced apoptosis through activating the JNK1/2-NF-κBp65 signaling pathway in AC16 cardiomyocytes; HMGB2 silencing inhibits I/R-induced JNK1/2 and NF-κBp65 activation; JNK1/2 inhibitor (SP600125) and NF-κB inhibitor (PDTC) reverse HMGB2 overexpression-induced injury. |
HMGB2 overexpression and shRNA knockdown, SP600125/PDTC pharmacological inhibition, western blot (JNK1/2, NF-κBp65, Bax/Bcl-2, caspase-3), CCK-8, flow cytometry, TUNEL |
Biomedicine & pharmacotherapy |
Medium |
30119172
|
| 2023 |
Post-translational modifications (PTMs) of HMGB2 are located predominantly in the B-domain and within the linker region (in contrast to HMGB1 where PTMs are in the A-domain and linker), as determined by MALDI mass spectrometry; despite high sequence homology, HMGB1 and HMGB2 have slightly different secondary structures as measured by CD spectroscopy. |
MALDI mass spectrometry (PTM mapping), UV circular dichroism spectroscopy |
International journal of molecular sciences |
Medium |
36834988
|
| 2023 |
HMGB2 is required for hepatic stellate cell (HSC) activation and liver fibrosis; Hmgb2-/- mice or HMGB2-inhibitor (inflachromene)-treated mice show impaired HSC transdifferentiation (reduced α-SMA) and slowed CCl4-induced fibrosis; AAV8-mediated HMGB2 overexpression enhances fibrosis; RNA-seq in Hmgb2-/- mice identifies top activated genes in integrin signaling, inflammation, cell cycle, and extracellular matrix pathways. |
Hmgb2-/- mice, AAV8-Hmgb2 overexpression, CCl4 fibrosis model, primary HSC isolation and transdifferentiation assay, small molecule inhibitor (inflachromene), lipo-shHMGB2, RNA-seq |
Hepatology communications |
High |
37930124
|
| 2016 |
HMGB2 promotes myocardial ischemic injury in rats by enhancing reactive oxygen species (ROS) production via RAGE signaling, thereby aggravating cardiomyocyte apoptosis, inflammation, and autophagosome clearance impairment; RAGE knockdown attenuates HMGB2-induced effects; TLR4 is not involved. |
Intramyocardial HMGB2 protein injection in MI rats, NADPH oxidase inhibitor (apocynin) co-treatment, RAGE knockdown (siRNA), ROS measurement, echocardiography, histopathology, western blot |
American journal of physiology. Heart and circulatory physiology |
Medium |
28011583
|
| 2018 |
HMGB2 knockdown in colorectal cancer cells reduces VSMC-like migration and invasion by stabilizing HMGB2 in the cytoplasm; lnc-CRCMSL physically binds HMGB2 and retains it in the cytoplasm, preventing nuclear entry; nuclear HMGB2 triggers EMT by interacting with OCT4; leptomycin B (nuclear export inhibitor) counteracts lnc-CRCMSL-mediated suppression by accumulating nuclear HMGB2. |
RNA pull-down assay (lnc-CRCMSL/HMGB2 interaction), subcellular fractionation, immunofluorescence, leptomycin B treatment, co-IP (HMGB2/OCT4), knockdown/overexpression, in vivo tumor model |
Oncogene |
High |
30575817
|
| 2020 |
miR-127-5p inhibits granulosa cell (GC) proliferation and impairs DNA damage repair capacity by targeting HMGB2; miR-127-5p upregulation in GCs from premature ovarian insufficiency patients reduces HMGB2 protein; in vivo, miR-127-5p attenuates DNA repair via HMGB2 in mouse ovary. |
miRNA mimic/inhibitor transfection, luciferase reporter assay (HMGB2 3'UTR), western blot, DNA damage repair assay, orthotopic mouse model, flow cytometry |
Journal of cellular physiology |
Medium |
32391592
|
| 1995 |
HMG2 mRNA level peaks at G2 phase of the cell cycle in rat fibroblasts; expression of antisense HMG2 RNA in COS-1 cells represses cell cycle progression at G1/S, resulting in decreased cell growth, indicating HMG2 expression is required for cell proliferation. |
Northern blot (cell cycle staging), antisense RNA expression (cell cycle analysis) |
Biochemical and biophysical research communications |
Medium |
7763232
|