Download Gene Section DNAJA3 (DnaJ (Hsp40) homolog, subfamily A, member 3)

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in Oncology and Haematology
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Gene Section
Review
DNAJA3 (DnaJ (Hsp40) homolog, subfamily A,
member 3)
June L Traicoff, Stephen M Hewitt, Joon-Yong Chung
Center for Peer Review and Science Management, SRA International, Inc Maryland, USA (JLT), Applied
Molecular Pathology Laboratory & Tissue Array Research Program, Laboratory of Pathology, Center for
Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA (SMH),
Applied Molecular Pathology Laboratory, Laboratory of Pathology, Center for Cancer Research, National
Cancer Institute, National Institutes of Health, Bethesda, MD, USA (JYC)
Published in Atlas Database: October 2011
Online updated version : http://AtlasGeneticsOncology.org/Genes/DNAJA3ID40342ch16p13.html
DOI: 10.4267/2042/47276
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence.
© 2012 Atlas of Genetics and Cytogenetics in Oncology and Haematology
Identity
DNA/RNA
Other names: FLJ45758, TID1, hTid-1
HGNC (Hugo): DNAJA3
Location: 16p13.3
Local order: According to NCBI Map Viewer, genes
flanking DNAJA3 are COR07-PAM16, NMRAL1, and
HMOX2.
Note
DNAJA3 was first identified by its ability to form
complexes with the human papillomavirus E7
oncoprotein (Schilling et al., 1998) in a yeast-two
hybrid screen. Sequence analysis revealed that
DNAJA3 was the human homolog of the Drosophila
tumor suppressor protein Tid56. Furthermore,
DNAJA3 contained a J-domain which is characteristic
of the family of DnaJ proteins which interact with and
stimulate the ATPase activity of heat shock cognate 70
(hsc70) family members (Schilling et al., 1998).
Note
DNAJA3 belongs to the evolutionarily conserved
DNAJ/HSP40 family of proteins. There are 41 known
DnaJ/Hsp40 proteins in the human genome (Qiu et al.,
2006).
According to NCBI Gene, the DNAJA3 gene is
conserved in human chimpanzee, cow, mouse, rat,
chicken, zebrafish, fruit fly, mosquito, C. elegans, S.
pombe, S. cerevisiae, K. lactis, E. gossypii, M. grisea,
N. crassa, and rice.
Description
The DNAJA3 gene is located on chromosome 16p13.3
between markers D16S521 and D16S418. This
chromosomal region carries several loci implicated in
human proliferation disorders, including the tuberous
sclerosis 2 gene (TSC2), polycystic kidney disease 1
gene (PKD1), and the CREB binding protein (CBP)
locus (Yin and Rozakis-Adcock, 2001).
Chromosome 16 - NC_000016.9. Modified from NCBI Map Viewer.
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DNAJA3 expression also increased in pathological
cardiac hypertrophic states (Hayashi et al., 2006).
DNAJA3 is approximately 34 kb and is composed of
12 exons separated by 11 introns. Exon sizes vary from
64 to 232 nucleotides, with the exception of exon 12
corresponding to the 3' untranslated region of
DNAJA3, which extends over 1.1 kb. Intron sizes vary
from 618 to 8291 nucleotides (Yin and RozakisAdcock, 2001).
Sequence encoding the DNAJ domain is present in
exons 2, 3 and 4, sequence encoding the Cys-rich
domain is found in exons 5 an 6, and the COOHterminal region is found in exons 7 through 11 (Yin and
Rozakis-Adcock, 2001).
Pseudogene
Paralogs. According to GeneCards, DNAJC16 is a
paralog for DNAJA3. DNAJC16 is located on
chromosome 1p36.1.
Protein
Note
The DNAJA3 gene encodes three cytosolic (Tid50,
Tid48, Tid46) proteins and three mitochondrial (Tid43,
Tid40, Tid38) proteins. Proteins encoded by the longer
splice variant DNAJA3L have often been designated in
the literature as Tid1L. Proteins encoded by the shorter
splice variant DNAJA3S have often been named Tid1S.
In this review, Tid1L will be designated DNAJA3L,
and Tid1S will be designated DNAJA3S. Specific
isoforms will be designated by size, e.g., Tid 50 will be
designated as DNAJA3 (50 kD).
Transcription
Promoter elements. DNAJA3 contains a putative
transcriptional start site 21 nucleotides upstream of the
initiating methionine. The presumptive promoter is
characterized by the lack of TATA and CAAT motifs,
and a high G+C content. The 5' flanking region
contains several consensus binding sites for
transcription factors that regulate gene expression
during tissue and organ development, such as myeloid
zinc finger (MZF1), Ikaros 2 and homeodomain
proteins, as well as factors implicated in cell growth
and survival responses, including AP-1, PEA3, E2F
and NF-kB.
Splice variants. Alternative splicing of a single
heteronuclear RNA (hnRNA) species generates the
three DNAJA3 isoforms. The long form DNAJA3L
(hTID1L) fully incorporates all exons. The
intermediate form DNAJA3I (hTID1I) is generated by
splicing of exon 10 to exon 12. This results in the loss
of the 34 C-terminal-most amino acids as well as the
stop codon; these are replaced with six amino acids
KRSTGN from exon 12. The short form DNAJA3S
(hTID1S) results from an in-frame deletion of 50 amino
acids that correspond precisely to exon 5 (Yin and
Rozakis-Adcock, 2001).
RNA expression. DNAJA3 mRNA was detected in 50
different human fetal and adult tissues. However the
relative abundance correlated with metabolic activity of
the tissues, with the highest levels observed in liver and
skeletal muscle (Kurzik-Dumke and Czaja, 2007).
Human tissues and cell lines showed differential
expression of the three DNAJA3 splice variant
mRNAs. Fetal brain tissue predominantly expressed
DNAJA3I, while breast tissues and T-cells
predominantly expressed DNAJA3L. Cell lines derived
from prostate epithelia, skin and lung fibroblasts,
normal astrocytes, and an osteosarcoma predominantly
expressed DNAJA3I with low levels of DNAJA3L also
present. DNAJA3S transcript was undetectable in all
samples (Yin and Rozakis-Adcock, 2001).
DNAJA3 transcripts showed differential expression
during development. Expression of DNAJA3
transcripts in mouse neonatal cardiomyocytes increased
as development of the heart proceeded and reached a
maximal level at 4 weeks of age, when cardiac
myocytes have matured (Hayashi et al., 2006).
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Description
DNAJA3 protein is present in two isoforms,
corresponding to splice variants encoding them. The
longer DNAJA3L isoform is a 480 amino acid protein
with a predicted size of 52 kD. The shorter DNAJA3S
isoform is a 453 amino acid protein with a predicted
size of 49 kD (Lu et al., 2006; UniProt).
Expression
DNAJA3 protein has been detected in human breast,
colon, ovarian, lung, and head and neck squamous cell
carcinoma (HNSCC) tissues (Traicoff et al., 2007;
Kurzik-Dumke et al., 2008; Chen et al., 2009).
Localisation
DNAJA3 localizes to human mitochondrial nucleoids,
which are large protein complexes bound to
mitochondrial DNA. Unlike other DnaJs, DNAJA3L
and DNAJA3S form heterocomplexes; both
unassembled and complexed DNAJA3 are observed in
human cells. DNAJA3L showed a longer residency
time in the cytosol prior to mitochondrial import as
compared with DNAJA3S; DNAJA3L was also
significantly more stable in the cytosol than DNAJA3S,
which is rapidly degraded (Lu et al., 2006).
Function
I. Binding partners
Human DNAJA3 protein has been shown to interact
with diverse partners, including viral proteins, heat
shock proteins, and key regulators of cell signaling and
growth.
Viral proteins
Hepatitis B virus core protein: DNAJA3 associated
with the hepatitis B virus core protein, specifically with
the carboxyl-terminal region (amino acids 94-185). The
N-terminal end of DNAJA3 (amino acids 1-447) was
required for this interaction. Furthermore, the
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terminus of DNAJA3L was required for this interaction
(Lu et al., 2006). Both DNAJA3S and DNAJA3L could
interact with Hsp70 (Kim et al., 2004). Endogenous
DNAJA3L and DNAJA3S coimmunoprecipitated with
mitochondrial Hsp70, but not Hsc70, in U2OS
osteosarcoma cells (Syken et al., 1999).
Tumor suppressor proteins
Adenomatous polyposis coli (APC): endogenous
cytosolic DNAJA3 proteins interacted with APC in
normal colon epithelium and colorectal cancer cell
lines (HT-29, Caco-2, and HRT-18). The N-terminal
Armadillo domain of APC was sufficient for binding to
DNAJA3. The DNAJA3 and APC interaction
comprised part of a larger multi-component complex
that also contained Hsp70, Hsc70, Actin, Dvl, and
Axin. This complex functions independently of the
known roles of APC in beta-catenin degradation and
proliferation mediated by Wg/Wnt signaling (KurzikDumke and Czaja, 2007).
Endogenous DNAJA3 proteins were shown to interact
with the caspase-cleaved N-terminus of APC in
HCT116 cells (Qian et al., 2010). The caspase-cleaved
APC protein has an important physiological role in
mediating apoptosis (Qian et al., 2010).
Patched: endogenous human Patched interacted with
the cytosolic forms of the DNAJA3 proteins in human
colon epithelium and colon tumor cells (Kurzik-Dumke
and Czaja, 2007). The tumor-associated polymorphism
in Patched (Ptch FVB allele) was associated with poorer
binding to DNAJA3 (Wakabayashi et al., 2007).
INT6: endogenous human DNAJA3 interacted with
INT6 (the p48 subunit of the eIF3 translation initiation
factor) in log phase, but not confluent, Jurkat T-cells
(Traicoff et al., 2007).
Von Hippel-Lindau protein (VHL): endogenous
pVHL co-immunoprecipitated with DNAJA3L protein
in HEK293 cells (Bae et al., 2005).
p53: DNAJA3 directly interacts with p53 through the
DNAJA3 DNAJ domain. Either the N- or C- terminal
domains of p53 was sufficient for the interaction (Trinh
et al., 2010).
Receptors
Interferon-gamma receptor (IFN-gammaR) subunit
IFN-gammaR2: DNAJA3 interacted with IFN-gamma
R2 in transfected COS cells. Furthermore, DNAJA3
bound more efficiently to a IFN-gammaR2 chimera
with an active kinase domain than to a similar construct
with an inactive kinase domain (Sarkar et al., 2001).
ErbB-2 (HER2/neu): endogenous ErbB-2 and
DNAJA3 co-immunoprecipitated in SK-BR-3 breast
cancer cells (Kim et al., 2004). The cytoplasmic
domains of ErbB-2 and DNAJA3 were sufficient for
this interaction (Kim et al., 2004).
ErbB-2 co-immunoprecipiated with DNAJA3 and the
carboxyl terminus of heat shock cognate 70 interacting
protein (CHIP). This complex was demonstrated in
tissue extracts from breast tumor specimens as well as
in transfected cell lines (Jan et al., 2011).
DNAJA3S precursor co-sedimented with viral capsidlike particles composed of the full-length core protein
(Sohn et al., 2006). Interaction between DNAJA3 and
the HBV core protein was confirmed in coimmunoprecipitation experiments using transfected
hepatoma cells (Sohn et al., 2006).
Epstein-Barr virus-encoded BARF1 protein:
DNAJA3 (amino acids 149-320) associated with the
Epstein-Barr virus-encoded BARF1 protein (amino
acids 21-221). Interaction between DNAJA3 and
BARF1 was confirmed in co-immunoprecipitation
experiments using transfected HeLa cells (Wang et al.,
2006).
Herpes simplex virus type 1 UL9 protein: DNAJA3
associated with the herpes simplex virus type 1 (HSV1) UL9 protein. UL9 protein is an origin-binding
protein. Interaction between DNAJA3 and UL9 was
confirmed by in vitro co-immunoprecipitation (Eom
and Lehman, 2002).
Human T cell leukemia virus type 1 (HTLV-1) Tax
protein: DNAJA3 associated with HTLV-1 Tax. The
interaction occurred through a central cysteine-rich zinc
finger-like region of DNAJA3 (amino acids 236 to
300). Interaction between DNAJA3 and Tax was
confirmed by co-immunoprecipitation experiments
using transfected human embryonic kidney cells (HEK)
(Cheng et al., 2001). Furthermore, the DNAJA3 and
Tax interaction occurred through a complex comprised
of DNAJA3, Tax, and heat shock protein 70 (Hsp70),
in which the cysteine-rich region of DNAJA3
interacted with Tax, while the J domain of DNAJA3
interacted with Hsp70 (Cheng et al., 2001).
Human
papilloma
virus-16
(HPV-16)
E7
oncoprotein: DNAJA3 was initially characterized
through its interaction with the HPV-16 E7
oncoprotein. DNAJA3 amino acids 1 to 235 and 297 to
342 independently interacted with HPV-16 E7.
Interaction between DNAJA3 and HPV-16 E7 was
confirmed by in vitro binding assays and coimmunoprecipitation experiments using transfected
human osteosarcoma (U2OS) cells (Schilling et al.,
1998).
Heat shock proteins
Hsp70 and Hsc70: endogenous DNAJA3 (specifically
the cytosolic form) immunoprecipitated with the heat
shock proteins Hsp70 and Hsc70 in normal colon
epithelium and colon cancer cell lines (Kurzik-Dumke
and Czaja, 2007). Endogenous DNAJA3 also interacted
with Hsp70/Hsc70 in HEp2 cells, and this interaction
was reduced in cells treated with interferon-gamma
(Sarkar et al., 2001). The J domain of DNAJA3 was
shown to be required for interaction with Hsp70 in
HEK cells (Cheng et al., 2001).
Proteins encoded by the long and short splice forms of
DNAJA3, DNAJA3L and DNAJA3S, respectively,
showed differential interactions with heat shock
proteins. Unassembled DNAJA3L (the long splice
variant) was shown to interact with Hsc70 specifically
in the cytosol (Lu et al., 2006). The unique carboxyl
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linking to the subsynaptic cytoskeleton, as
demonstrated by knockdown and overexpression
experiments.
Knockdown of DNAJA3 in skeletal muscle fibers
resulted in dispersed synaptic AchR clusters and
impaired neuromuscular transmission. Knockdown of
DNAJA3 in myotubes resulted in inhibition of AchR
clustering, inhibition of agrin-induced activation of the
Rac and Rho small GTPases and tyrosine
phosphorylation of AchR, and decreased Dok-7induced clustering of AChRs. In contrast,
overexpression of the N-terminal half of DNAJA3
induced agrin-and MuSK-independent phosphorylation
and clustering of AChRs (Linnoila et al., 2008; Song
and Balice-Gordon, 2008).
Neurite outgrowth: DNAJA3 regulated nerve growth
factor (NGF)-induced neurite outgrowth in PC12derived nnr5 cells. DNAJA3 bound to Trk at the
activation loop and DNAJA3 was tyrosine
phosphorylated by Trk in yeast cells, transfected cells,
and in neurotophin-stimulated primary rat hippocampal
neurons. Overexpression of DNAJA3 led to NGFinduced neurite outgrowth in TrkA-expressing nnr5
cells. In contrast, knockdown of DNAJA3 resulted in
reduced NGF-induced neurite growth in nnr5-TrkA
cells (Liu et al., 2005).
Viral pathways
Hepatitis B virus replication: expression of DNAJA3
suppressed replication of HBV in human hepatoma
cells, while knockdown of DNAJA3 led to increased
HBV replication. The mechanism for inhibited
replication was through accelerated degradation and
destabilization of the viral core and HBx proteins (Sohn
et al., 2006).
Herpes simplex virus type 1 replication: the HSV-1
UL9 protein is an origin-binding protein that is
essential for viral DNA replication. DNAJA3
modulates DNA replication by enhancing the binding
of UL9 protein to an HSV-1 origin and facilitating
formation of the multimer from the dimeric UL9
protein, perhaps through a chaperone function.
However, DNAJA3 had no effect on the DNAdependent ATPase or helicase activities associated with
the UL9 protein (Eom and Lehman, 2002).
Epstein-Barr virus secretion: the EBV BARF1 gene
encodes a secretory protein with transforming and
mitogenic activities. Coexpression experiments with
BARF1 and DNAJA3 showed that DNAJA3 could
promote secretion of BARF1, perhaps through
chaperone functions (Wang et al., 2006).
Motility and metastasis
DNAJA3 was shown to negatively regulate the motility
and metastasis of breast cancer cells through
attenuation of nuclear factor kappaB activity on the
promoter of the IL8 gene (Kim et al., 2005).
Reductions of DNAJA3 levels in MDA-MB231 breast
cancer cells increased their migration as a result of
increased interleukin-8 (IL-8) secretion without
affecting survival or growth rate. Furthermore,
Trk receptor tyrosine kinases: the carboxyl-terminal
end of DNAJA3 (residues 224-429) bound to Trk at its
activation loop in a phosphotyrosine-dependent manner
(Liu et al., 2005).
Muscle-specific kinase (MuSK) component of the
agrin receptor: DNAJA3S, but not DNAJA3L,
associated with the cytoplasmic portion of MuSK in
mouse skeletal muscle cells (Linnoila et al., 2008).
Signaling proteins
NF-kappaB: DNAJA3 strongly associated with the
cytoplasmic protein complex of NF-kappaB-IkappaB
through
direct
interaction
with
IkappaBalpha/IkappaBbeta and the IKKalpha/beta
subunits of the IkappaB kinase complex. The
endogenous interaction was observed in Jurkat, SAOS2, and HEK293 cells (Cheng et al., 2005).
JAK/STAT: Jak2 interacted with DNAJA3S as well as
DNAJA3L as shown by immunoprecipitation from
transfected COS-1 cells expressing these proteins.
Endogenous DNAJA3 and Jak2 were shown to interact
in HEp2 cells (Sarkar et al., 2001).
The carboxyl terminus of endogenous DNAJA3L, but
not DNAJA3S, co-immunoprecipitated with STAT1
and with STAT3 in U2OS cells (Lu et al., 2006).
DNAJA3L remained associated with activated
phosphorylated STAT1 upon treatment with interferongamma (Lu et al., 2006).
The DNAJ domain of DNAJA3 interacted with the
transactivation domain of Stat5b in hematopoietic cell
lines (Dhennin-Duthille et al., 2011).
p120 GTPase-activating protein (GAP): both the
cytoplasmic precursor and mitochondrial mature forms
of murine DNAJA3 associated with GAP in vivo in
rodent cells. GAP selectively bound to the
unphosphorylated form of murine DNAJA3 (Trentin et
al., 2001).
DNA replication proteins
DNA polymerase gamma (Polga) alpha subunit:
endogenous DNAJA3 interacted with the alpha subunit
of Polga in HEK293 cells. Polga is the only
mitochondrial DNA polymerase responsible for all
mitochondrial DNA synthetic reactions (Hayashi et al.,
2006).
II. Signaling pathways and cellular effects
DNAJA3 modulates diverse signaling pathways and
cellular effects that are vital for cell growth and
differentiation.
Neural pathways
Neuromuscular synaptogenesis: DNAJA3 is an
essential component of the agrin signaling pathway that
is crucial for synaptic development. Motoneuronderived agrin clusters nicotinic acetylcholine receptors
(AChRs) in mammalian cells. DNAJA3 binds to the
cytoplasmic domain of muscle-specific kinase (MuSK),
a component of the agrin receptor and colocalizes with
AchRs at developing, adult, and denvervated motor
endplates. DNAJA3 transduces signals from MuSK
activation to AchR clustering, culmintating in cross-
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regardless of the p53 expression status. In contrast,
cells transduced with a DNAJA3 mutant that has an Nterminal J domain deletion and that lost suppressive
activity on IKK, continued to proliferate (Cheng et al.,
2005).
DNAJA3 mediates apoptosis through the Bcl-2
pathway. DNAJA3 induced apoptosis in SF767 glioma
cells that contained a tumor-associated mutation at the
DNAJA3 locus. Apoptosis resulted from caspase
activation and cytochrome c release from mitochondria.
However, Bcl-XL protected cells from hTid-1Sinduced cell death and cytochrome c release. However,
hTid1S caused S and G2/M arrest in cells with wild
type Tid1. Interestingly, hTid1L had no effect on
growth of glioma cells (Trentin et al., 2004).
Immature dnaja3(-/-) DN4 thymocytes exhibited
significantly reduced expression of the antiapoptotic
bcl-2 gene (Lo et al., 2005).
Expression of constitutively active AKT (pAKT)
counteracted and inhibited DNAJA3-induced apoptosis
in HNSCC cells (Chen et al., 2009).
DNAJA3 mediates apoptosis through APC.
DNAJA3 (40 kD) isoform inhibited apoptosis through
antagonizing the apoptotic function of the N-terminal
region of the APC protein (Qian et al., 2010).
DNAJA3 mediates apoptosis through p53.
Overexpression of DNAJA3 enhanced p53-dependent
apoptosis, and restored pro-apoptotic activity of mutant
p53 in colon, breast, and glioma cell lines (Ahn et al.,
2010). The mechanism is through direct interaction of
the DNAJ domain of DNAJA3 and p53 (Trinh et al.,
2010). In contrast, depletion of DNAJA3 resulted in the
inhibition of hypoxia or genotoxic stress-induced p53
mitochondrial localization and apoptosis (Trinh et al.,
2010).
Mitochondrial functions
Although DNAJA3 has many cellular functions,
DNAJA3 often localizes to the mitochondria and also
has important functions in the mitochondria.
Epidermal growth factor (EGF) response: GAP and
DNAJA3 were shown to colocalize at perinuclear
mitochondrial membranes in response to EGF
stimulation (Trentin et al., 2001).
p53 localization and apoptotic function: depletion of
DNAJA3 prevented p53 accumulation at the
mitochondria and resulted in resistance to apoptosis
under hypoxic or genotoxic stresses (Trinh et al.,
2010). DNAJA3 formed a complex with p53 under
hypoxic conditions that directed p53 translocation to
the mitochondria and the subsequent initiation of
apoptosis (Ahn et al., 2010). Loss of DNAJA3
expression abrogated p53 translocation to the
mitochondria and inhibited apoptosis (Ahn et al.,
2010). Conversely, overexpression of DNAJA3
promoted p53 mitochondrial localization and apoptosis
(Ahn et al., 2010).
Viral protein localization: in the absence of Tax,
expression of the DNAJA3/Hsp70 molecular complex
was targeted to perinuclear mitochondrial clusters. In
DNAJA3 was shown to negatively modulate de novo
synthesis of IL-8 through regulating NFkappaB
activity. Additionally, DNAJA3 knockdown enhanced
the metastasis of breast cancer cells in animals (Kim et
al., 2005).
Other studies also indicate a potential role for DNAJA3
in inhibiting transformation and metastasis. Stable
DNAJA3 knockdown cells exhibited an enhanced
ability for anchorage-independent growth, as measured
by an increase in soft-agar colony formation (Edwards
and Münger, 2004). In contrast, ectopic expression of
DNAJA3 in HNSCC cells was shown to significantly
inhibit cell proliferation, migration, invasion,
anchorage-independent
growth,
and
xenotransplantation tumorigenicity (Chen et al., 2009).
Expression of DNAJA3 inhibited the transformation
phenotype of two human lung adenocarcinoma cell
lines (Cheng et al., 2001).
Apoptosis
DNAJA3 encodes two mitochondrial matrix localized
splice variants: DNAJA3 (43 kD) and DNAJA3 (40
kD). DNAJA3 (43 kD) and DNAJA3 (40 kD) do not
themselves induce apoptosis; instead they have
opposing effects on apoptosis induced by exogenous
stimuli.
Expression of DNAJA3 (43 kD) increases apoptosis
induced by both the DNA-damaging agent mitomycin c
and tumor necrosis factor-alpha. This activity is J
domain-dependent, since a J domain mutant of
DNAJA3 (43 kD) suppressed apoptosis. Conversely,
expression of DNAJA3 (40 kD) suppressed apoptosis,
while expression of a J domain mutant of DNAJA3 (40
kD) increased apoptosis (Syken et al., 1999).
Cells lacking expression of DNAJA3 proteins were
protected from cell death in response to multiple
stimuli, including cisplatin, tumor necrosis factor
alpha/cycloheximide and mitomycin C (Edwards and
Münger, 2004).
DNAJA3 regulates activation-induced cell death
(AICD) in the Th2 subset of helper T cells. AICD is an
apoptotic process induced by stimulation through the
T-cell receptor and Th2 cells are significantly less
prone to AICD than Th1 cells are. The antiapoptotic
variant, Tid-1S was shown to be selectively induced in
murine Th2 cells following activation. Expression of a
dominant-negative mutant of hTid-1S in a Th2 cell line
strikingly enhanced activation of caspase 3 in response
to CD3 stimulation, and caused the cells to become
sensitive to AICD. Therefore, the accumulation of Tid1S in Th2 cells following activation may contribute to
the induction of apoptosis resistance during the
activation of Th2 cells (Syken et al., 2003).
DNAJA3 mediates apoptosis through the nuclear
factor kappaB (NF-kappaB) pathway. DNAJA3
repressed the activity of NF-kappaB through physical
and functional interactions with the IKK complex and
IkappaB. Overexpression of DNAJA3 led to inhibition
of cell proliferation and induction of apoptosis of
human osteosarcoma cells and human melanoma cells
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fibroblasts, as well as in premature senescence of
REF52 cells triggered by activated ras. Conversely,
spontaneous immortalization of rat embryo fibroblasts
was suppressed upon ectopic expression of DNAJA3.
Suppression of endogenous DNAJA3 activity
alleviated the suppression of tumor necrosis factor
alpha-induced NF-kappaB activity by DNAJA3. These
results suggest that DNAJA3 contributes to senescence
by repressing NF-kappa B signaling (Tarunina et al.,
2004).
DNAJA3 repressed NF-kappaB activity induced by
Tax, tumor necrosis factor alpha (TNFalpha), and
Bcl10. DNAJA3 specifically suppressed serine
phosphorylation of IkappaBalpha by activated IkappaB
kinase beta (IKKbeta).
The suppressive activity of DNAJA3 on IKKbeta
required a functional J domain that mediates
association with heat shock proteins and resulted in
prolonging the half-life of the NF-kappaB inhibitors
IkappaBalpha and IkappaBbeta (Cheng et al., 2002).
AKT: overexpression of DNAJA3 in HNSCC cells
inhibited cell proliferation, migration, invasion,
anchorage-independent
growth,
and
xenotransplantation tumorigenicity. Overexpression of
DNAJA3 attenuated EGFR activity and blocked the
activation of AKT in HNSCC cells, which are known
to be involved in the regulation of survival in HNSCC
cells. Conversely, ectopic expression of constitutively
active AKT greatly reduced apoptosis induced by
DNAJA3 overexpression (Chen et al., 2009).
JAK2: DNAJA3L and DNAJA3S interacted with Jak2
in vivo in COS-1 cells. Interaction was primarily in the
cytoplasm and predominantly with the active kinase
domain of Jak2 (Sarkar et al., 2001).
c-MET receptor tyrosine kinase (MetR): MetR
interacted with DNAJA3L and DNAJA3S, but showed
preferential binding to DNAJA3S. Interaction occurred
through the J domain. In RCC cells, overexpression of
DNAJA3S
enhanced
HGF-mediated
MetR
autophosphorylation, while DNAJA3L showed modest
inhibition of MetR activity. Modulation of MetR
phosphorylation levels was independent of pVHL.
DNAJA3S enhanced HGF-mediated cell migration and
modulated HGF-mediated MAPK phosphorylation.
DNAJA3 knockdown inhibited MetR activation and
migration in response to HGF (Copeland et al., 2011).
Signal transducers and activators of transcription
(STAT) 5b: DNAJA3 specifically interacted with
STAT5b but not STAT5a in hematopoietic cell lines.
Interaction involved the DNAJ domain. DNAJA3
negatively regulated the expression and transcriptional
activity of STAT5b and suppressed the growth of
hematopoietic cells transformed by an oncogenic form
of STAT5b (Dhennin-Duthille et al., 2011).
Cell Fate
DNAJA3 was shown to be required for the T-cell
transition from double-negative 3 to double-positive
stages. Mice with dnaja3 specifically deleted in T cells
developed thymic atrophy, with dramatic reduction of
the presence of Tax, DNAJA3 and its associated Hsp70
are sequestered within a cytoplasmic "hot spot"
structure, a subcellular distribution that is characteristic
of Tax in HEK cells (Cheng et al., 2001).
APC interaction: the amino terminus of APC
interacted with DNAJA3 at the mitochondria in vivo in
colorectal cancer cell lines (Qian et al., 2010).
Chaperone function: DNAJA3 isoforms were also
shown to exhibit a conserved mitochondrial DnaJ-like
function substituting for the yeast mitochondrial DnaJlike protein Mdj1p (Lu et al., 2006).
Mitochondrial biogenesis: DNAJA3 was shown to be
crucial for mitochondrial biogenesis partly through
chaperone activity on DNA polymerase gamma
(Hayashi et al., 2006). Mice deficient in Dnaja3
developed dilated cardiomyopathy (DCM) and died
before 10 weeks of age (Hayashi et al., 2006).
Progressive respiratory chain deficiency and decreased
copy number of mitochondrial DNA were observed in
cardiomyocytes lacking Dnaja3 (Hayashi et al., 2006).
Tumor suppressor pathways
APC: DNAJA3 directly bound to the APC tumor
suppressor protein and promoted a physiological
function for APC that was independent of APC's
involvement in beta-catenin degradation or regulation
of the actin cytoskeleton (Kurzik-Dumke and Czaja,
2007).
pVHL: TID1L directly interacted with von HippelLindau protein and enhanced the interaction between
HIF-1 alpha and pVHL. This resulted in destabilization
of HIF-1 alpha protein, decreased vascular endothelial
growth factor expression, and inhibition of
angiogenesis (Bae et al., 2005).
Interferon-gamma: DNAJA3L and DNAJA3S
interacted with the interferon-gamma receptor chain
IFN-gammaR2 and modulated IFN-gamma-mediated
transcriptional activity. Furthermore, IFN-gamma
treatment
reduced
the
interaction
between
Hsp70/Hsc70 and DNAJA3 (Sarkar et al., 2001).
Oncogenic pathways
Erb-B2/HER2: DNAJA3 physically interacted with
the signaling domain of ErbB-2 and ErbB-2 were
shown to colocalize in mammary carcinoma cells (SKBR-3). Overexpression of DNAJA3 induced growth
arrest and apoptosis in ErbB-2-overexpressing breast
cancer cells; the DNAJ and C-terminal domains of
DNAJA3 were critical for mediating apoptosis.
Downregulation of ERK1/ERK2 and BMK1 MAPK
pathways also contributed to apoptosis. DNAJA3S
negatively regulated ErbB-2 signaling pathways by
enhancing the degradation of ErbB-2. Finally,
increased cellular DNAJA3 inhibited the growth of
ErbB-2-dependent tumors in mice (Kim et al., 2004).
Mammary tumor tissue from breast cancer patients and
transgenic mice carrying the rat HER-2/neu oncogene
suggest that DNAJA3 suppresses ErbB-2 in breast
cancers (Kurzik-Dumke et al., 2010).
NF-kappaB: expression of DNAJA3 was upregulated
upon cellular senescence in rat and mouse embryo
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DNAJA3 (DnaJ (Hsp40) homolog, subfamily A, member 3)
Traicoff JL, et al.
correlation with negative or weakly positive expression
of ErbB2 in human breast cancer tissue samples. High
DNAJA3 levels were strongly correlated with high
levels of CHIP (carboxyl terminus of heat shock
cognate 70 interacting protein). Lower expression of
DNAJA3 had a higher risk of unfavorable tumor grade,
later pathological stage, larger tumor size, and
microscopic features of a more malignant histology
(Jan et al., 2011). Higher expression of DNAJA3
correlated with increased 10-year overall and diseasefree survival rate (Jan et al., 2011).
The expression of the three DNAJA3 isoform
transcripts was examined in human breast cancer
carcinomas by RT-PCR. Aberrant expression of all
three forms correlated with malignant transformation.
Furthermore, elevated DNAJA3L expression was
associated with less aggressive tumors (Kurzik-Dumke
et al., 2010). Immunohistochemical analysis
demonstrated high levels of DNAJA3 protein in tumors
of the luminal A subtype, but significantly lower levels
of DNAJA3 protein in the luminal B subtype, triple
negative tumors, and the HER-2 subtype which
overexpresses HER-2 (Kurzik-Dumke et al., 2010).
Multiplex tissue immunoblotting of human breast
tumor tissue microarrays was used to test correlations
between DNAJA3 protein levels and a set of tumor
suppressor proteins. DNAJA3 protein levels showed
strongly positive correlations with p53, Patched, and
INT6 proteins (Traicoff et al., 2007). Additionally,
DNAJA3 protein levels showed moderate positive
correlations with c-Jun and phospho-c-Jun proteins
(Traicoff et al., 2007).
double-positive and single-positive thymocytes in the
dnaja3(-/-) thymus. DNAJA3 deficiency inhibited cell
proliferation and enhanced cell death of DN4 cells. The
expression profile of genes involved in cytokine
receptor signaling was altered in DN4 T-cells.
Expression of human bcl-2 transgene restored T
lymphocyte proliferation and differentiation in the
dnaja3 knockout mice. These results suggest that
dnaja3 is critical in early thymocyte development,
especially during transition from the DN3 to doublepositive stages, possibly through its regulation of bcl-2
expression, which provides survival signals.
Homology
Mouse (laboratory): Dnaja3
Rat: Dnaja3
Cattle: DNAJA3
Chimpanzee: DNAJA3
Dog (domestic): DNAJA3
Mutations
Note
The SF767 glioma cell line exhibits an aberrant 52 kD
molecular weight protein. Sequence analysis of cDNA
generated from this line showed two mutations: an
additional thymine at nucleotide position 1438 and an
additional cytosine at nucleotide position 1449. These
mutations alter the reading frame of the DNAJA3
sequence, introducing an additional 71 amino acids
following the penultimate threonine residue at position
479. The mutations appear to increase the steady-state
abundance of the mutant protein, resulting in aberrantly
high levels of the DNAJA3 mutant variant (Trentin et
al., 2004).
Head and neck squamous cell
carcinoma (HNSCC)
Disease
The clinical association between DNAJA3 expression
and progression of HNSCC was explored using
immunohistochemical analysis of primary HNSCC
patient tumor tissue. DNAJA3 expression was
negatively associated with tumor T stage, overall stage,
survival, and recurrence. Patients with higher
expression of DNAJA3 were predicted to have better
overall survival than those with low or undetectable
expression of DNAJA3 protein (Chen et al., 2009).
Highly malignant HNSCC cell lines also demonstrated
low or undetectable levels of DNAJA3, in contrast to
less aggressive lines where DNAJA3 protein was easily
detected (Chen et al., 2009).
Implicated in
Colon cancer
Disease
DNAJA3 and INT6 protein levels, as well as DNAJA3
and Patched protein levels, were positively correlated
in human colon tumor tissues (Traicoff et al., 2007).
However, there were no correlations between DNAJA3
and p53, c-Jun, or phospho-c-Jun protein levels
(Traicoff et al., 2007). These results were demonstrated
by multiplex tissue immunoblotting of tissue
microarrays (Traicoff et al., 2007).
Progression of colorectal cancers correlated with
overexpression and loss of polarization of expression of
DNAJA3. These changes were associated with
upregulation
of
Hsp70
and
loss
of
compartmentalization of APC (Kurzik-Dumke et al.,
2008).
Ovarian cancer
Disease
Multiplex tissue immunoblotting of ovarian tumor
tissues demonstrated that DNAJA3 protein levels
showed moderate positive correlations with INT6, cJun, phospho-c-Jun, and p53. No correlations were
observed between DNAJA3 and Patched (Traicoff et
al., 2007).
Breast cancer
Disease
DNAJA3 protein expression showed a strong
Atlas Genet Cytogenet Oncol Haematol. 2012; 16(3)
202
DNAJA3 (DnaJ (Hsp40) homolog, subfamily A, member 3)
Traicoff JL, et al.
Edwards KM, Münger K. Depletion of physiological levels of
the human TID1 protein renders cancer cell lines resistant to
apoptosis mediated by multiple exogenous stimuli. Oncogene.
2004 Nov 4;23(52):8419-31
Lung cancer
Disease
Multiplex tissue immunoblotting of lung tumor tissues
demonstrated that DNAJA3 protein levels were
strongly correlated with INT6. DNAJA3 protein levels
were moderately correlated with Patched, c-Jun, and
p53. However, DNAJA3 proteins showed negative
correlation with phospho-c-Jun in these samples
(Traicoff et al., 2007).
Kim SW, Chao TH, Xiang R, Lo JF, Campbell MJ, Fearns C,
Lee JD. Tid1, the human homologue of a Drosophila tumor
suppressor, reduces the malignant activity of ErbB-2 in
carcinoma cells. Cancer Res. 2004 Nov 1;64(21):7732-9
Tarunina M, Alger L, Chu G, Munger K, Gudkov A, Jat PS.
Functional genetic screen for genes involved in senescence:
role of Tid1, a homologue of the Drosophila tumor suppressor
l(2)tid, in senescence and cell survival. Mol Cell Biol. 2004
Dec;24(24):10792-801
Cardiomyopathy
Note
Mice deficient in Dnaja3 developed dilated
cardiomyopathy (DCM) and died before 10 weeks of
age (Hayashi et al., 2006). Progressive respiratory
chain deficiency and decreased copy number of
mitochondrial DNA were observed in cardiomyocytes
lacking Dnaja3 (Hayashi et al., 2006).
Trentin GA, He Y, Wu DC, Tang D, Rozakis-Adcock M.
Identification of a hTid-1 mutation which sensitizes gliomas to
apoptosis. FEBS Lett. 2004 Dec 17;578(3):323-30
Bae MK, Jeong JW, Kim SH, Kim SY, Kang HJ, Kim DM, Bae
SK, Yun I, Trentin GA, Rozakis-Adcock M, Kim KW. Tid-1
interacts with the von Hippel-Lindau protein and modulates
angiogenesis by destabilization of HIF-1alpha. Cancer Res.
2005 Apr 1;65(7):2520-5
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Traicoff JL, Hewitt SM, Chung JY. DNAJA3 (DnaJ (Hsp40)
homolog, subfamily A, member 3). Atlas Genet Cytogenet
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