| 1997 |
HAX-1 was identified as a novel intracellular protein that directly associates with HS1 (a substrate of Src family tyrosine kinases) via the amino-terminal region of HS1 and the carboxyl-terminal half of HAX-1; HAX-1 localizes predominantly to mitochondria but also to endoplasmic reticulum and nuclear envelope. |
Yeast two-hybrid screening, co-immunoprecipitation, confocal microscopy, deletion mutant analysis |
Journal of immunology |
High |
9058808
|
| 2000 |
HAX-1 interacts with the polycystic kidney disease protein PKD2 (but not the closely related PKD2L) and also associates with the F-actin-binding protein cortactin, linking PKD2 to the actin cytoskeleton; PKD2 and HAX-1 colocalize in the cell body and in cellular processes and lamellipodia. |
Yeast two-hybrid screen, immunofluorescence colocalization, specificity demonstrated by failure of PKD2L to interact |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
10760273
|
| 2002 |
KSHV K15 protein interacts with HAX-1 via its C-terminus both in vitro and in vivo; HAX-1 colocalizes with K15 at the ER and mitochondria; HAX-1 can form homodimers in vivo and functions as a potent inhibitor of apoptosis. |
Yeast two-hybrid screen, in vitro and in vivo binding assays, immunofluorescence colocalization, apoptosis assays |
Journal of virology |
Medium |
11752170
|
| 2002 |
HAX-1 binds specifically to the 3' untranslated region of vimentin mRNA; HAX-1 and eEF-1gamma form protein complexes that interact with vimentin 3'UTR and can be pulled from HeLa cell extracts using a vimentin 3'UTR RNA affinity column, establishing HAX-1 as an RNA-binding protein. |
Yeast three-hybrid assay, in vitro RNA-protein binding, RNA affinity pulldown from cell extracts |
Nucleic acids research |
Medium |
12466525
|
| 2004 |
HAX-1 is a specific substrate of the mitochondrial serine protease Omi/HtrA2; Omi cleaves HAX-1 both in vitro and in vivo during apoptosis; HAX-1 degradation is an early event occurring while Omi is still confined to mitochondria; inhibition of Omi protease activity prevents HAX-1 degradation and reduces cell death. |
In vitro cleavage assay, in vivo cleavage in cells treated with apoptotic stimuli, Omi-specific inhibitor, cell line with catalytically inactive Omi (mnd2 mice-derived) |
The Journal of biological chemistry |
High |
15371414
|
| 2004 |
Gα13 physically interacts with HAX-1, and this interaction is required for Gα13-stimulated cell migration; HAX-1 expression reduces actin stress fibers and focal adhesion complexes in Gα13-expressing cells; HAX-1 attenuates Gα13-stimulated RhoA activity while potentiating Rac activity; a quaternary complex of Gα13, HAX-1, Rac, and cortactin was identified; siRNA-mediated silencing of HAX-1 drastically reduces Gα13-mediated cell migration. |
Co-immunoprecipitation, siRNA knockdown, cell migration assay, RhoA/Rac activity assays, actin/focal adhesion staining |
The Journal of biological chemistry |
High |
15339924
|
| 2004 |
HAX-1 binds to BSEP, MDR1, and MDR2 ABC transporters; HAX-1 co-localizes with BSEP and MDR1 at the apical membrane of MDCK cells; RNAi-mediated depletion of HAX-1 increases BSEP levels in the apical membrane by 71% by enhancing retention (not by altering synthesis, modification, or delivery); HAX-1 also interacts with cortactin and participates in BSEP internalization from the apical membrane. |
Yeast two-hybrid, GST pulldown, co-immunoprecipitation, RNA interference, pulse-chase labeling, subcellular fractionation, immunofluorescence |
The Journal of biological chemistry |
High |
15159385
|
| 2006 |
HAX-1 is critical for maintaining the inner mitochondrial membrane potential in myeloid cells; loss-of-function HAX1 mutations (homozygous) cause increased apoptosis in myeloid cells and severe congenital neutropenia. |
Positional cloning, germline mutation identification, mitochondrial membrane potential assay, apoptosis assay in patient myeloid cells |
Nature genetics |
High |
17187068
|
| 2006 |
HAX-1 interacts with caspase-9 and inhibits caspase-9 processing in a dose-dependent manner in a cell-free caspase activation assay; HAX-1 overexpression in adult cardiac myocytes protects against apoptosis; on apoptotic stimulation, caspase-9 translocates to mitochondria and colocalizes with HAX-1. |
Yeast two-hybrid, cell-free caspase activation assay with recombinant protein, overexpression/siRNA knockdown in cardiac myocytes, immunofluorescence colocalization |
Circulation research |
High |
16857965
|
| 2006 |
HAX-1 interacts with phospholamban (PLN) via PLN residues 16–22 and HAX-1 residues 203–245; binding affinity ~1 µM measured by surface plasmon resonance; phosphorylation of PLN by PKA reduces HAX-1 binding; Ca2+ diminishes PLN/HAX-1 interaction dose-dependently; HAX-1 redistributes from mitochondria to ER upon co-transfection with PLN; PLN enhances HAX-1 anti-apoptotic effects against hypoxia/reoxygenation. |
Yeast two-hybrid, GST pulldown, surface plasmon resonance, co-transfection/colocalization, apoptosis assay |
Journal of molecular biology |
High |
17241641
|
| 2007 |
HAX-1 binds the 3'UTR hairpin motif of DNA polymerase beta mRNA exclusively as a dimer; disruption of the hairpin impairs RNA-protein complex formation; HAX-1 was detected in the nuclear matrix in addition to mitochondria, consistent with a role in post-transcriptional regulation. |
In vitro RNA binding assay, luciferase reporter system, mutagenesis of hairpin, subcellular fractionation |
Nucleic acids research |
Medium |
17704138
|
| 2008 |
HAX-1 interacts with the mitochondrial proteases PARL and HtrA2/Omi; HAX-1 presents HtrA2 to PARL, facilitating proteolytic processing of HtrA2 to the active form in the mitochondrial intermembrane space; processed HtrA2 prevents accumulation of mitochondrial-outer-membrane-associated activated Bax, blocking apoptosis; Hax1 deficiency in mice causes apoptosis in lymphocytes and neurons. |
Genetic (Hax1 knockout mouse), co-immunoprecipitation, epistasis with Parl and HtrA2 mutants, Bax localization assay, apoptosis assay |
Nature |
High |
18288109
|
| 2008 |
HAX-1 also binds to SERCA2 via SERCA2 residues 575–594 interacting with the C-terminal domain of HAX-1 (aa 203–245); HAX-1 overexpression down-regulates SERCA2 protein levels and reduces ER Ca2+ levels; SERCA2 overexpression abrogates the protective effects of HAX-1 on cell survival after hypoxia/reoxygenation; PLN co-transfection causes massive redistribution of HAX-1 to ER where it co-distributes with PLN and SERCA2. |
Deletion mapping, co-immunoprecipitation, cell transfection/colocalization, cell viability assay, Ca2+ measurement |
Molecular biology of the cell |
High |
18971376
|
| 2008 |
HAX1 mutations affecting both transcript variants 1 and 2 cause severe congenital neutropenia with neurological symptoms (epilepsy, neurodevelopmental delay), while mutations affecting only transcript variant 1 cause neutropenia without neurological symptoms; transcript variant 2 is markedly expressed in human brain tissue. |
Mutation screening of SCN patients, genotype-phenotype analysis, tissue expression analysis of HAX1 isoforms |
Blood |
Medium |
18337561
|
| 2008 |
HIV-1 Vpr physically associates with HAX-1; Vpr overexpression dislocates HAX-1 from mitochondria and causes mitochondrial instability and cell death; HAX-1 overexpression suppresses Vpr's pro-apoptotic activity. |
Co-immunoprecipitation, overexpression studies, mitochondrial localization assay, cell viability assay |
Journal of virology |
Medium |
16227293
|
| 2009 |
HAX-1 overexpression reduces SERCA2a pump activity in isolated cardiomyocytes and in vivo, depressing calcium kinetics and contractility; conversely, HAX-1 downregulation enhances calcium cycling; HAX-1 promotes formation of PLN monomers (the active/inhibitory units of SERCA2a); ablation of PLN rescues HAX-1 inhibition of contractility in vivo, placing HAX-1 upstream of PLN in cardiac calcium regulation. |
Cardiac-specific overexpression mouse model, HAX-1 knockdown, calcium kinetics measurement, contractility assays, PLN monomer/pentamer analysis, genetic rescue (PLN ablation) |
Proceedings of the National Academy of Sciences of the United States of America |
High |
19920172
|
| 2009 |
HAX-1 variant I and II localize selectively to mitochondrial membranes, while variants III, IV, and V localize to both mitochondria and sarcoplasmic reticulum; deletion of HAX-1's NH2-terminus abolishes mitochondrial targeting and attenuates anti-apoptotic capacity; removal of the PLN-binding site prevents HAX-1 translocation to SR; HAX-1 is preferentially lost from SR of PLN-deficient hearts. |
Immunoelectron microscopy, subcellular fractionation, deletion analysis, cellular transfection, colocalization, PLN-deficient mouse hearts |
Journal of molecular and cellular cardiology |
High |
19913549
|
| 2009 |
HAX-1 in vivo does not interact with PARL because the two proteins are confined in distinct cellular compartments; sequence analysis shows HAX-1 lacks authentic Bcl-2 homology (BH) modules and is unlikely to be a Bcl-2 family protein; in vitro interaction between Hax1 and PARL is proposed to be an artifact. |
In vivo localization/fractionation, sequence analysis and secondary structure prediction |
Cell death and differentiation |
Low |
19680265
|
| 2010 |
Granzyme B inserts into mitochondria and cleaves HAX-1 into two fragments: an N-terminal fragment that remains in mitochondria and a C-terminal fragment released to cytosol; the N-terminal fragment acts as a dominant negative, causing mitochondrial depolarization in a cyclophilin D-dependent manner; overexpression of wild-type HAX-1 or an uncleavable mutant protects against GrB-mediated depolarization. |
In vitro cleavage assay, mitochondrial fractionation, overexpression of cleavage mutants, membrane potential measurement, cyclophilin D dependence assay |
The Journal of biological chemistry |
High |
20388708
|
| 2010 |
HAX-1 interacts with XIAP via HAX-1 C-terminal domain binding to XIAP BIR2 and BIR3 domains (each with affinity similar to full-length XIAP by SPR); HAX-1 suppresses polyubiquitination of XIAP, enhancing its stability and thereby inhibiting apoptosis. |
Proteomic screen (immunoprecipitation + 2D gel), GST pulldown, surface plasmon resonance, ubiquitination assay, cell viability assay |
Biochemical and biophysical research communications |
Medium |
20171186
|
| 2011 |
Hax1 depletion in neutrophil-like PLB-985 cells impairs uropod detachment and directed migration; Hax1-deficient cells show increased integrin-mediated adhesion and reduced RhoA activity; RhoA depletion phenocopies Hax1 loss; activation of RhoA rescues adhesion defects in Hax1-deficient cells, placing Hax1 upstream of RhoA in regulating neutrophil adhesion and chemotaxis. |
siRNA knockdown, microfluidic migration assay, integrin-mediated adhesion assay, RhoA activity assay, epistasis (RhoA activation rescue) |
The Journal of cell biology |
High |
21518791
|
| 2012 |
HAX-1 is a nucleocytoplasmic shuttling protein dependent on exportin 1 (CRM1/XPO1); two nuclear export signals (NES) were identified by systematic mutagenesis; nuclear accumulation occurs after leptomycin B treatment or specific cellular stress; HAX-1 status influences mRNA levels of DNA polymerase beta (one of its mRNA targets); HAX-1 tethering to reporter transcript decreases its expression; HAX-1 colocalizes with P-body markers. |
Mutagenesis of NES, leptomycin B treatment, nuclear/cytoplasmic fractionation, reporter assay, P-body colocalization |
The FEBS journal |
Medium |
23164465
|
| 2012 |
HAX1 interacts with influenza A virus PA polymerase subunit via the nuclear localization signal (NLS) domain of PA; HAX1 knockdown increases nuclear accumulation of PA and enhances viral polymerase activity and virus yield, which can be reversed by HAX1 re-expression; thus HAX1 impedes nuclear transport of PA and restricts influenza A virus propagation. |
Yeast two-hybrid, GST pulldown, co-immunoprecipitation, HAX1 knockdown and rescue, nuclear fractionation, minigenome assay, virus yield assay |
Journal of virology |
High |
23055567
|
| 2012 |
HCLS1 (HS1) is phosphorylated upon G-CSF stimulation and binds LEF-1, transporting it to the nucleus; HAX1 mutations in SCN patients impair G-CSF-triggered HCLS1 phosphorylation and reduce LEF-1 autoregulation, establishing HAX1 as required for HCLS1 phosphorylation in G-CSF signaling and granulopoiesis. |
Phosphorylation assays, co-immunoprecipitation, nuclear fractionation, patient myeloid cells, HCLS1-deficient mouse (neutropenic phenotype) |
Nature medicine |
High |
23001182
|
| 2012 |
HAX-1 is rapidly degraded by the proteasome via K48-linked ubiquitination dependent on its PEST sequence; a PEST-deletion mutant of HAX-1 is more resistant to proteasomal degradation and exerts greater anti-apoptotic protection than wild-type HAX-1. |
PEST sequence deletion mutagenesis, ubiquitination assay, proteasome inhibitor treatment, apoptosis assay |
BMC cell biology |
Medium |
22827267
|
| 2012 |
HAX-1 overexpression inhibits the IRE-1 ER stress signaling pathway by binding to the N-terminal fragment of Hsp90; HAX-1 sequesters Hsp90 away from IRE-1 to the PLN-SERCA2a complex; loss of HAX-1 in heterozygous-deficient hearts increases infarct size and IRE-1 activity; the Hsp90 inhibitor 17-AAG abolishes HAX-1's IRE-1 inhibitory effects. |
Cardiac overexpression mouse model, HAX-1 heterozygous-deficient mouse, co-immunoprecipitation (HAX-1/Hsp90), pharmacological (17-AAG), ischemia/reperfusion model |
Circulation research |
High |
22982986
|
| 2013 |
HAX-1 variant 1 (v1) is anti-apoptotic while rat v2/human v4 is pro-apoptotic; Hax-1 isoforms form homotypic and heterotypic dimers with binding affinities ranging from ~3.8 nM (v1 homodimers) to ~97 nM (v1/v2 heterodimers); the minimal dimerization region spans aa 97-278; co-expression of v1 and v2 neutralizes both pro- and anti-apoptotic activities through modulation of cytochrome c release. |
SPR binding kinetics, deletion analysis, overexpression in epithelial cells, cytochrome c release assay, myocardial infarction model |
The Journal of biological chemistry |
High |
24347163
|
| 2014 |
PRKCD (PKCδ) phosphorylates both FBXO25 and HAX-1, directing nuclear FBXO25 to mitochondrial HAX-1; FBXO25 is the substrate-recognition subunit of SCF(FBXO25) ubiquitin ligase that ubiquitinates and degrades HAX-1 after apoptotic stress; stabilizing HAX-1 phosphodegron mutations found in human MCL inhibit apoptosis; FBXO25 re-expression in FBXO25-deleted MCL cells promotes cell death. |
Unbiased substrate screen, co-immunoprecipitation, ubiquitination assay, phosphorylation assay, FBXO25 re-expression in MCL cells, Eµ-Myc mouse lymphoma model, xenotransplant model |
Nature medicine |
High |
25419709
|
| 2014 |
Hax-1 is required for Rac1-cortactin interaction and colocalization in ovarian cancer cells; Hax-1 interacts with cortactin via domains aa 1-56 (Hax-D1) and aa 113-168 (Hax-D3), and with Rac1 via domains aa 57-112 (Hax-D2) and aa 169-224 (Hax-D4); silencing of Hax-1 reduces LPA-stimulated migration of SKOV3 cells and impairs Rac1-cortactin colocalization; expression of individual Hax-1 domains competitively inhibits migration. |
siRNA knockdown, domain mapping by deletion/truncation, co-immunoprecipitation, colocalization, cell migration assay |
Genes & cancer |
Medium |
25053987
|
| 2015 |
HAX-1 regulates cyclophilin-D (CypD) protein levels via an Hsp90-dependent mechanism: HAX-1 overexpression interferes with CypD binding to Hsp90, rendering CypD susceptible to ubiquitin-proteasomal degradation; HAX-1 overexpression enhances CypD ubiquitination; reduced CypD decreases mPTP activation; proteasome inhibition or elevated Hsp90 rescues CypD levels in HAX-1-overexpressing cells. |
Cardiac-specific HAX-1 overexpression and heterozygous KO mouse models, co-immunoprecipitation, ubiquitination assay, proteasome inhibitor, genetic ablation of CypD, mPTP opening assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
26553996
|
| 2015 |
Hax-1 interacts with microtubule end-binding protein EB2 in an EB2-specific manner; knockdown of either HAX1 or EB2 in skin epidermal cells stabilizes focal adhesions and impairs epidermal migration in vitro and in vivo; cell motility and focal adhesion turnover require the Hax1-EB2 interaction. |
Quantitative proteomics (EB2 interactome), co-immunoprecipitation, siRNA knockdown, focal adhesion assay, in vitro and in vivo migration assay |
The Journal of biological chemistry |
Medium |
26527684
|
| 2016 |
Kv3.3 (KCNC3) recruits Arp2/3 to the plasma membrane via binding of its cytoplasmic C-terminus to Hax-1, forming a stable cortical actin network that prevents rapid N-type channel inactivation; a human disease-causing Kv3.3 mutation within the conserved proline-rich domain binds Hax-1 but fails to recruit Arp2/3, resulting in growth cones with deficient actin veils in stem cell-derived neurons. |
Co-immunoprecipitation, cytochalasin D resistance assay, electrophysiology, stem cell-derived neurons, disease mutation analysis |
Cell |
High |
26997484
|
| 2017 |
HAX-1 ablation in the adult heart (cardiac-specific inducible KO) increases calcium affinity of SERCA2a and reduces PLN-SERCA2a binding, demonstrating that endogenous HAX-1 mediates approximately 50% of PLN's inhibitory activity on cardiac calcium cycling and contractility; PLN overexpression in HAX-1-null cardiomyocytes has no inhibitory effect, indicating HAX-1 limits PLN activity. |
Cardiac-specific inducible HAX-1 knockout mouse, calcium kinetics, SERCA2a Ca2+ affinity measurement, PLN-SERCA2a co-immunoprecipitation, isoproterenol stimulation, PLN overexpression rescue |
The Journal of biological chemistry |
High |
29150445
|
| 2017 |
HAX-1 ablation in the adult heart causes increased reactive oxygen species production at the SR/ER compartment, leading to SERCA2a oxidation and enhanced proteolysis; HAX-1 interacts with NADPH oxidase 4 (NOX4), a newly identified binding partner; NOX4 inhibition abrogates the detrimental effects of HAX-1 ablation in ischemia/reperfusion injury. |
Cardiac-specific inducible HAX-1 KO mouse, ROS measurement, SERCA2a oxidation assay, co-immunoprecipitation (HAX-1/NOX4), NOX inhibitor (apocynin) |
Journal of molecular and cellular cardiology |
High |
29169992
|
| 2018 |
H5N1 PB1-F2 binds to HAX-1 (a host restriction factor of IAV PA); the PA subunit of mammal-adapted H1N1 is resistant to HAX-1 restriction while avian-origin H5N1 PA remains sensitive; PB1-F2 alleviates HAX-1's inhibition of H5N1 polymerase activity through direct competition with HAX-1 for PA binding. |
Co-immunoprecipitation, polymerase activity assay, siRNA/overexpression of HAX-1, virus replication assay with PB1-F2-deficient mutant |
Journal of virology |
Medium |
29563290
|
| 2019 |
HAX1 knockdown affects actomyosin contractility through changes in RhoA and septin signaling, impairing collective (but not single-cell) migration, cell-cell junctions, and cell layer integrity in breast cancer cells. |
HAX1 siRNA knockdown, collective/single-cell migration assays, RhoA activity assay, septin staining, cell-cell junction analysis |
Molecular biology of the cell |
Medium |
31644363
|
| 2020 |
Hepatic HAX-1 interacts with inositol 1,4,5-trisphosphate receptor-1 (InsP3R1); HAX-1 absence reduces InsP3R1 levels, improving ER-mitochondria calcium homeostasis to prevent excess mitochondrial calcium overload; HAX-1 ablation activates pyruvate dehydrogenase and increases mitochondrial utilization of glucose and fatty acids; HAX-1 deficiency also increases bile salt exporter protein (BSEP) levels, promoting enterohepatic bile acid recirculation. |
Liver-specific HAX-1 KO mouse, co-immunoprecipitation, InsP3R1 quantification, calcium assays, metabolic phenotyping, pyruvate dehydrogenase activity |
The Journal of biological chemistry |
High |
32079675
|
| 2021 |
Kv3.3 channels bind and stimulate TBK1 (TANK-binding kinase 1); TBK1 activity is required for Kv3.3 binding to its auxiliary subunit Hax-1 (which prevents channel inactivation); a disease-causing Kv3.3 mutation overactivates TBK1, leading to Hax-1 accumulation in multivesicular bodies and lysosomes, loss of Hax-1, caspase activation, and neuronal cell death. |
Co-immunoprecipitation, TBK1 kinase assay, dominant negative/inhibitor studies, subcellular trafficking assays, electrophysiology, caspase activity assay |
Nature communications |
High |
33741962
|
| 2022 |
HAX1 and CLPB control the balance of mitochondrial protein synthesis and persistence (mitochondrial proteostasis) as shown by SILAC proteomics; impaired mitochondrial protein dynamics in HAX1-deficient cells are associated with decreased abundance of PRKD2 and phosphorylated HSP27 (Ser78/82); cellular defects in HAX1-/- cells can be functionally reconstituted by HSP27, defining a CLPB/HAX1/(PRKD2)/HSP27 axis critical for neutrophil granulocyte differentiation. |
SILAC proteomics, HAX1-deficient cells/mouse models, PRKD2 and HSP27 quantification, functional reconstitution with HSP27 |
The Journal of clinical investigation |
High |
35499078
|
| 2024 |
EIF3H functions as a deubiquitinase for HAX1, stabilizing HAX1 by antagonizing βTrCP-mediated ubiquitination; stabilized HAX1 enhances the interaction among RAF1, MEK1, and ERK1, potentiating ERK1/2 phosphorylation and promoting colorectal cancer progression. |
Co-immunoprecipitation, ubiquitination assay, deubiquitination assay, conditional Eif3h deletion mouse model, RAF1-MEK1-ERK1 interaction assay, patient-derived xenografts |
Nature communications |
High |
38514606
|