European Journal of Cell Biology 93 (2014) 98105

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European Journal of Cell Biology

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Mini review

Emerging roles of MCPH1: Expedition from primary

microcephaly to cancer
Thejaswini Venkatesh a, , Padmanaban S. Suresh b
a
Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, Karnataka, India
b
Centre for Biomedical Research, Vellore Institute of Technology University, Vellore, Tamil Nadu, India

a r t i c l e i n f o a b s t r a c t

Article history: Genetic mutations in microcephalin1 (MCPH1) cause primary autosomal recessive microcephaly which
Received 19 October 2013 is characterized by a marked reduction in brain size. MCPH1 encodes a centrosomal protein with three
Received in revised form 9 January 2014 BRCT (BRCA1 C-terminal) domains. Also, it is a key regulator of DNA repair pathway and cell cycle check-
Accepted 15 January 2014
points. Interestingly, in the past few years, many research studies have explored the role of MCPH1, a
neurodevelopmental gene in several cancers and its tumor suppressor functions have been elucidated.
Keywords:
Given the diverse new emerging roles, it becomes critical to review and summarize the multiple roles of
MCPH1
MCPH1 that is currently lacking in the literature. In this review after systematic analysis of literature, we
BRIT1
Autosomal recessive primary microcephaly
summarise the multiple functional roles of MCPH1 in centrosomal, DNA repair and apoptotic pathways.
DNA repair Additionally, we discuss the considerable efforts taken to understand the implications of MCPH1 in dis-
Cancer eases such as primary microcephaly and its other emerging association with cancer and otitis media. The
Otitis media promising view is that MCPH1 has distinct roles and its clinical associations in various diseases makes it
Apoptosis an attractive therapeutic target.
Glioblastoma 2014 Elsevier GmbH. All rights reserved.
OSCC
miR-27a

Introduction
The rare genetically heterogenous autosomal recessive pri-
mary microcephaly, also called as MCPH (Micro-Cephaly Primary
Hereditary), is characterized by the clinical features such as OFC
(occipital frontal circumference) of less than 3 SDs below the
mean for age and sex, mild to moderate mental retardation and,
with no further neurological ndings except for mild seizures
(Passemard et al., 2009; Kaindl et al., 2010). However, the true
phenotype spectrum of MCPH patients is wider than indicated pre-
viously and now, includes a simplied gyral pattern, periventricular
neuronal heterotopias (abnormal positioning of the neurons in
the brain), polymicrogyria (development of numerous microgyri),
speech delay, hyperactivity and attention decit, aggressiveness,
focal or generalized seizures, and delay of developmental mile-
stones and pyramidal signs (Kaindl et al., 2010). Totally, mutations
in thirteen genes have been identied to cause MCPH. They
include MCPH1 (microcephalin 1), WDR62 (WD repeat domain
62), CDK5RAP2 (cyclin-dependent kinase 5 regulatory associated
Corresponding author at: Department of Molecular Reproduction, Development
and Genetics, Indian Institute of Science, Bangalore 560012, India.
Tel.: +91 9663068681.
E-mail address: thejaswinivenkatesh5@gmail.com (T. Venkatesh).
protein 2), CASC5 (cancer susceptibility candidate 5), CEP152 (cen-
trosomal protein 152 kDa), ASPM (asp [abnormal spindle] homolog,
microcephaly associated [Drosophila]), CENPJ (centromeric protein
J), STIL (SCL/TAL1-interrupting locus), CEP135 (centrosomal pro-
tein 135 kDa), CEP63 (centrosomal protein 63 kDa), ZNF335 (zinc
nger protein 335), PHC1 (polyhomeotic homolog 1 [Drosophila])
and MKL2 (MKL/myocardin-like 2) (Bond et al., 2002; Jackson et al.,
2002; Bond et al., 2005; Kumar et al., 2009; Guernsey et al., 2010;
Nicholas et al., 2010; Sir et al., 2011; Hussain et al., 2013; Yang et al.,
2012; Awad et al., 2013; Ramos et al., 2013). Among these genes,
MCPH1 was the rst gene to be identied as a causative gene for
MCPH (Jackson et al., 1998; Jackson et al., 2002). It was mapped to
chromosome 8p22-pter using autozygosity mapping in two con-
sanguineous Pakistani families. Jackson et al. (1998) identied a
minimal region of 13.2 cM, dened by the markers D8S1824 and
D8S1825D8S and D8S1825 and subsequently, identied MCPH1 as
the causative gene. In this study, both the consanguineous families
showed a single homozygous mutation c.74C>G(p.Ser25X) in the
second exon of MCPH1(Jackson et al., 2002). Also, another homozy-
gous missense mutation c.80C>G(p.Tyr27Arg) in a MCPH patient
of Caucasian origin was identied (Trimborn et al., 2005). Eight
mutations have been described for MCPH1 in a study consisting
of 112 Iranian families with MCPH (Darvish et al., 2010). Addi-
tionally, three more mutations in MCPH1 have been reported in
MCPH patients (Farooq et al., 2010; Ghani-Kakhki et al., 2012;

0171-9335/$ see front matter 2014 Elsevier GmbH. All rights reserved.
http://dx.doi.org/10.1016/j.ejcb.2014.01.005
 T. Venkatesh, P.S. Suresh / European Journal of Cell Biology 93 (2014) 98105
Hussain et al., 2013). In addition to MCPH, it is interesting to note
that mutations in MCPH1 have been associated with other disease
conditions where microcephaly is one of the distinct features. A
homozygous mutation c.427 428insA(p.Thr143fsX5) in MCPH1 has
been reported in PCC (premature chromosome condensation) syn-
drome which is characterized by microcephaly and postnatal short
stature. PCC is a phenomenon where mitotic chromosomes con-
dense at the interphase stage itself (Neitzel et al., 2002; Trimborn
et al., 2004). Moreover, a deletion of 150200 kb encompassing
the promoter and rst 6 exons of MCPH1 has been reported in an
Iranian consanguineous family with autosomal recessive mental
retardation (ARMR) and mild microcephaly (Garshasbi et al., 2006).
Although it is widely studied in relation to primary microcephaly,
recent studies suggest correlation of MCPH1 mutations and its dys-
functions with cancer and otitis media (OM), and this has further
strengthened its implications in different tissues. Its identication
and clinical correlation has also suggested that they have diverse
roles apart from neurodevelopment. Given the important roles of
MCPH1 and its emerging correlations with cancer biology and other
diseases, a greater understanding of its role is undeniably needed.
In this review, considering the recent advances, we highlight its
structure, expression and localization, targets and functions, and
recent insights into its role in cancer development and OM.
Structure
The MCPH1 gene has 14 coding exons (Jackson et al., 2002).
Recently, four alternative transcripts of human MCPH1 have been
identied in 562T broblast cells. The isoforms include: full-
length (FL) and the alternative transcripts 914 (914 exons are
deleted), 13 (13 exons are deleted), and 8 (exon 8 is deleted)
(Gavvovidis et al., 2012). The human MCPH1 protein is 835 amino
acids long and shows 57% sequence identity with the mouse pro-
tein of 822 amino acids (Jackson et al., 2002; Kumar et al., 2002). It
consists of three BRCT (BRCA1 C-terminal) domains which extend
from amino acids 7 to 83 (BRCT1), 642 to 720 (BRCT2) and 753 to
823 (BRCT3) (Jackson et al., 2002). BRCT2 and BRCT3 constitute the
C-terminal twin BRCT domains. BRCT domains are a new class of
phosphoprotein binding modules that are found in the components
of DNA repair pathways (Huyton et al., 2000; Glover et al., 2004).
MCPH1 protein has a NLS (nuclear localization signal) from 355 to
375 amino acids, and deletion of amino acid residues 301400 of
MCPH1 leads to the detainment of MCPH1 within the cytoplasm
of HEK293T (human embryonic kidney cells) (Wood et al., 2007).
The evolutionary signicance of the MCPH1 protein has been stud-
ied. The MCPH1 protein from 11 species (7 primates and 4 other
mammalian species) were aligned to gain insights into the evolu-
tionary signicance of the protein (Shi et al., 2013). The amino acid
residues (M96, S101, V310, H314, T377, Y425, L442, R485 and P835)
of the human protein were conserved across the species. However,
the amino acid residues (I161, E167, A510 and S841) occurred in
the ancestor of hominidae and hence, these are great-ape specic
residues (Shi et al.., 2013).
Expression and localization
MCPH1 is ubiquitously expressed in adult human tissues, and
its highest expression is observed in the brain, testes, pancreas and
liver (Lin et al., 2010). Its transcripts are highly expressed in human
fetal brain, kidney and liver, and expressed less in human fetal heart
and lungs (Jackson et al., 2002). Mcph1 has been detected in the
lateral ventricles of the developing forebrain of the fetal mouse
brain (Jackson et al., 2002). Additionally, MCPH1 transcripts have
been detected at all developmental stages from embryos to adults
in Drosophila (Brunk et al., 2007).
99
MCPH1 localizes to centrosomes in U2OS (osteosarcoma) and
DT-40 (chicken B) cells (Zhong et al., 2006; Jeffers et al., 2008). It also
localizes to DNA repair foci in response to DNA damage in U2OS,
HeLa (cervical carcinoma), and microcephalic patient derived lym-
phoblastoid cells (Rai et al., 2006; Wood et al., 2007; Gavvovidis
et al., 2010). In addition to cell lines, the localization of MCPH1
has been studied in human tissue sections. A study has shown that
MCPH1 localizes to nuclear foci, the nucleus and the cytoplasm in
primary cultures of normal ovarian tissues (Brning-Richardson
et al., 2011). A study of IHC (immunohistochemistry) reported
that MCPH1 localization was conned to the nucleus and nuclear
foci of normal breast tissues (Richardson et al., 2011). On a more
physiological scenario, localization of MCPH1 has been studied in
Drosophila. MCPH1 localizes to the interphase DNA in syncytial
embryos of Drosophila, and it is absent on the DNA of cells in the
prometaphase to telophase stages of Drosophila syncytial embryos
(Brunk et al., 2007).
Phenomics of MCPH1 model organisms
Drosophila as a model organism
The full-length MCPH1 gene in Drosophila is designated as
MCPH1 (L). The MCPH1 (L) protein contains an N-terminal and two
tandem C-terminal BRCT domains. MCPH1 (L) undergoes alterna-
tive splicing to give three more isoforms. The short isoform, MCPH1
(S) contains the N-terminal BRCT domain. The other two isoforms
are short of 47 amino acids at their N-terminal ends and are sim-
ilar to MCPH1(L) and MCPH1(S) isoforms (Brunk et al., 2007). The
MCPH1 transcripts (S and L forms) show highest expression at the
syncitial and gastrula stages and, lowest expression at the blas-
tula stage of the y development. MCPH1 protein is expressed at
a higher level in ovaries and a lower level in the brain of the lar-
vae (Brunk et al., 2007). Furthermore, two Drosophila lines with
the homozygous deletion of either 13 or 14 exons have been
generated (Brunk et al., 2007). These mutant ies were viable and
showed no abnormalities of the brain. In contrast to this study, Rick-
myre et al. (2007) reported that the MCPH1 mutant homozygous
male ies showed defective brain structures ranging from mal-
formed lobes to complete absence of the lobes, though the sizes
of the brain were unaffected. But, the MCPH1 mutant homozy-
gous female ies showed no defects in the size or the structures
of the brain (Rickmyre et al., 2007). Also, the mutant ies showed
no morphological or behavioural defects in adults (Brunk et al.,
2007). However, the embyros laid by the MCPH1 null female ies
lacked cellularization and showed defective synticial embryogen-
esis (Brunk et al., 2007).
Mouse as a model organism
Few groups have elucidated the functions of MCPH1 in mouse
models. A mouse model with impaired Mcph1 function (referred
as Mcph1gt/gt mouse) was generated using the gene trap method
(Trimborn et al., 2010). Trimborn et al. (2010) introduced a gene
trap in intron 12 of Mcph1, which led to the deletion of C-
terminal BRCT domain. In this study, the body and brain sizes
of Mcph1gt/gt mice were within the range of wild type controls.
However, the overall survival rates of Mcph1gt/gt mice were sig-
nicantly reduced compared to wild type and heterozygous mice.
No signs of premature malignant disease or impaired fertility were
reported in Mcph1gt/gt mice (Trimborn et al., 2010). Surprisingly,
the cell cultures derived from th Mcph1gt/gt mice appeared to be
largely normal after irradiation and showed no signs of defective
DNA damage responses (Trimborn et al., 2010). However, the pri-
mary broblast cultures of the homozygous Mcph1 mutant mice
100 T. Venkatesh, P.S. Suresh / European Journal of Cell Biology 93 (2014) 98105
exhibited PCC. Also, the authors claim that these discrepancies of
the Mcph1 functions in Mcph1gt/gt mice is due to the residual wild
type Mcph1 expression which was indeed detected by the quan-
titative real-time RT-PCR in the spleen, brain and liver (Trimborn
et al., 2010). Liang et al. (2010) generated a Mcph1 global knock-
out mouse by deleting the exon 2 using the cre/lox system. These
Mcph1/ mice survived to adulthood, but were growth-retarded
and infertile (Liang et al., 2010). However, the Mcph1/ mice did
not develop any tumors over a period of one and a half years. Mouse
embryonic broblasts (MEFs) and T lymphocytes derived from
these Mcph1/ mice were hypersensitive to -irradiation (Liang
et al., 2010). Furthermore, a failure to carry out meiotic recombi-
nation was observed in spermatocytes, and this failure has been
attributed to the phenomenon of infertility seen in these knockout
mice (Liang et al., 2010). Another Mcph1/ mice model was gener-
ated by the disruption of the Mcph1 gene using the cre/lox system
(Gruber et al., 2011). Their study demonstrated that the Mcph1-
del (Mcph1 knockout) mice showed reduced brain size and less
neuroprogenitor cell proliferation. Gruber et al. (2011) developed
neurospheres from both the Mcph1-del and Mcph1-ctr (littermate
controls) mice and reported that the number and size of the neu-
rospheres were not affected in Mcph1-del mice as compared to
that of the Mcph1-ctr mice. However, the tendency to form sec-
ondary neurospheres was decreased in the Mcph1-del mice when
compared to the neurospheres derived from the Mcph1-ctr mice.
Cells of the neurospheres derived from Mcph1-del mice underwent
premature mitotic entry, which eventually lead to the uncoupling
of the mitosis and the centrosomal cycle, leading to misorienta-
tion of the mitotic spindles (Gruber et al., 2011). Misorientation of
mitotic spindles alters the division plane, leading to a premature
shift from symmetric to asymmetric divisions in the neuropro-
genitor cells (Gruber et al., 2011). Thus, the increased asymmetric
divisions in neuroprogenitor cells led to the reduction in the num-
ber of neurons and microcephaly. Also, these mice did not exhibit
tumor development over a span of 2 years (Gruber et al., 2011).
Additionally, Zhou et al. (2013) have examined whether the Mcph1
deletion would affect DNA damage response in neural cells of
Mcph1-del mice. The absence of Mcph1 compromised HR in Mcph1-
del neuroprogenitors (Zhou et al., 2013). The authors discuss that
the inefcient HR could cause DNA damage accumulation in cells
leading to genomic instability and nally apoptosis. Corroborating
this concept, increased apoptosis has been observed in Mcph1-del
neuroprogenitors (Gruber et al., 2011; Zhou et al., 2013). Further-
more, increased apoptosis may also contribute to neuronal loss in
addition to shift in asymmetric divisions during the development
of microcephaly. Hence, the studies on MCPH1 in model organisms
have lead to a better understanding of its functional roles in the
development of MCPH. However, there are no studies that report
Mcph1 overexpressing mouse models. There is a need for future
studies on MCPH1 transgenic mice to enhance our understanding
of the protein function.
MCPH1 targets and functions
Centrosomal functions of MCPH1
MCPH1 localizes to centrosomes in U2OS and DT-40 cells, and
the N-terminal BRCT domain mediates its centrosome localization
(Zhong et al., 2006; Jeffers et al., 2008; Rai et al., 2008). MCPH1
depleted DT-40 cells displayed normal barrel structures of the cen-
trioles (Brown et al., 2010). However, MCPH1 deciency lead to
IR induced centrosome amplication in DT-40, U2OS and human
MCPH1 mutant lymphoblastoid cells (Alderton et al., 2006; Rai et al.,
2008; Brown et al., 2010). It is surprising to note that the centro-
some amplication seen in DT-40 cells after exposure to IR was
abrogated with the disruption of Pcnt (Wang et al., 2013). MCPH1
null blood lymphocytes derived from patients with MCPH and PCC
syndrome exhibit PCC (Trimborn et al., 2004; Trimborn et al., 2005).
PCC is caused by the lack of a functional N-terminal BRCT domain of
MCPH1 (Wood et al., 2008; Richards et al., 2010; Yamashita et al.,
2011). Also, centrosomal function of MCPH1 has been associated
with PCC. MCPH1 interacts with PCNT (pericentrin) and directs
CHEK1 (checkpoint kinase 1) to centrosomes (Tibelius et al., 2009).
CHEK1 mediates the inhibitory phosphorylation of centrosomal
CDK1 (cyclin-dependent kinase 1) at Y15. CDK1 and CCNB1 (cyclin
B1) accumulate in the centrosome during interphase, and activa-
tion of the CDK1-CCNB1 complex is the key event to enter into
mitosis (Nurse, 1990; Bailly et al., 1992). The centrosomal activa-
tion of CDK1 and CCNB1 occurs in the late prophase (Jackman et al.,
2003). Regulation of centrosomal CDK1 and CCNB1 is very critical
to synchronise the cytoplasmic and nuclear mitotic events. In the
absence of MCPH1 in MCPH1 patients, the recruitment of CHEK1
to the centrosomes by MCPH1 is restricted and hence, CDK1 is
not inhibited by CHEK1. The MCPH1 mutant PCC cells exhibit stri-
kingly low levels of pY15-CDK1 (Alderton et al., 2006). This leads
to the early activation of the CCNB1-CDK1 complex and hence,
mitosis starts prematurely leading to PCC (Tibelius et al., 2009).
Additionally, CDC25 phosphatases regulate CDK1-CCNB1 activity
by reversing inhibitory Y15 phosphorylation on CDK1, and the low
levels of pY15-CDK1 levels were attributed to lack of CHEK1 medi-
ated proteosomal degradation of CDC25 phosphatases (Alderton
et al., 2006). Thus, MCPH1 regulates mitotic entry by regulating the
activities of CDC25A and CDK1-CCNB1 (Alderton et al., 2006).
Additionally, another non-centrosomal mechanism has been
proposed to account for the feature of PCC seen in the MCPH1
depleted condition. It has been demonstrated that the N-terminal
BRCT domain of MCPH1 interacts with the SET (SET nuclear onco-
gene) protein and MCPH1/ broblast cells depleted of SET by
siRNA exhibited profound PCC (Leung et al., 2011). But, it is still not
known how the MCPH1-SET complex generates PCC. However, it
is known that SET prevents histone acetylation and regulates chro-
matin compaction (Miyamoto et al., 2003). Hence, MCPH1 could be
aiding chromatin compaction functions of SET protein.
DNA repair functions of MCPH1
The new DNA repair functional roles of MCPH1 are reshaping
the current understanding of the DNA repair pathways. MCPH1
functions as a proximal factor for the DNA damage repair path-
way and regulates the ATM (ataxia telangiectasia mutated) and ATR
(ataxia telangiectasia and Rad3 related) pathways (Rai et al., 2006).
In response to DNA damage, MCPH1 co-localizes with numerous
proteins such as -H2AX (H2A histone family, member X), MDC1
(mediator of DNA-damage checkpoint 1), TP53BP1 (tumor protein
p53 binding protein 1), NLRP2 (NLR family, pyrin domain contain-
ing 2), p-ATM, ATR and p-RAD17 [RAD17 homolog (S. pombe)] to
DNA damage repair foci in U2OS cells (Fig. 1) (Rai et al., 2006). H2AX
is the proximal signalling protein, and upon DNA damage, it gets
phosphorylated at Y142 and S139 by WSTF (Williams-Beuren syn-
drome transcription factor), and ATM or ATR kinases, respectively.
MCPH1 functions as a versatile sensor of H2AX phosphorylation
marks by recognizing both of these phosphorylation tags (Wood
et al., 2007; Shao et al., 2012; Singh et al., 2012a). Also, the C-
terminal BRCT domain of MCPH1 interacts with the N-terminal
region of BRCA2 (breast cancer 2, early onset) and positively regu-
lates the localization of BRCA2 to DNA damage repair foci (Wu et al.,
2009). Moreover, MCPH1 interacts with ATM/ATR phosphorylated
SMARCC2 (SWI/SNF related, matrix associated, actin dependent
regulator of chromatin, subfamily c, member 2), which is a sub-
unit of the SWI-SNF complex upon DNA damage. This interaction
of MCPH1 and SWI/SNF complex leads to chromatin relaxation
 T. Venkatesh, P.S. Suresh / European Journal of Cell Biology 93 (2014) 98105 101

Fig. 1. Functions of MCPH1. MCPH1 is represented by the central green box. The protein/complex interacting with MCPH1 is shown as boxes/other shapes in contact with
it. The transcriptional activation and inactivation are indicated by vertical red and green arrows respectively. The six major functions of MCPH1 such as apoptosis, DNA
repair, centrosomal integrity, telomere maintenance, chromatin relaxation and cell cycle checkpoints are represented in each purple box. (A) MCPH1 mediates apoptosis
by upregulation of pro-apoptotic genes in E2F1 dependent manner. (B) MCPH1 interacts with -H2AX and BRCA2 for the formation of ionizing radiation induced foci (IRIF)
indicated within the dashed boxes. (C) MCPH1 maintains the centrosome number and spindle alignment. (D) It interacts with the telosome and maintains telomere integrity.
It also transcriptionally represses the activity of hTERT. (E) MCPH1 interacts with SWI/SNF complex, relaxes the chromatin and increases the accessibility to DNA repair
proteins. (F) It regulates G1/S and G2/M cell cycle checkpoints by transcriptional activation of BRCA1 and CHEK1. The diagram is based on various studies cited appropriately in
the text.(For interpretation of the references to colour in this gure legend, the reader is referred to the web version of this article.)

(opening of the DNA) and allows the recruitment of DNA repair with MCPH1 using its [Y/F] XL domain and maintains the telomere
proteins to the sites of DNA damage (Peng et al., 2009). Further- integrity in HTC75 (human brosarcoma) cells (Kim et al., 2009).
rmore, U2OS cells depleted of MCPH1 by siRNAs are defective in
HR (homologous recombination) and NHEJ (non-homologous end
MCPH1 represses hTERT
joining) pathways (Peng et al., 2009). Two more studies have con-
rmed the role of MCPH1 in HR using DR-GFP/I-sceI system in
MCPH1 was initially identied as a repressor of hTERT in an
MCPH1/ MEFs and Mcph1-del ES cells (Wood et al., 2008; Zhou
ERM (enhanced retroviral mutagens) screen in HeLa cells (Lin
et al., 2013). Furthermore, MCPH1 interacts with Condensin II, a
and Elledge, 2003). In support of the hTERT repressing activity of
protein complex required for chromosome condensation and seg-
MCPH1, it has been shown that MCPH1 binds to the proximal region
regation. But surprisingly, it has been shown that the interaction of
of the hTERT promoter and represses its transcription in HeLa cells
MCPH1 with Condensin II mediates HR and does not have any chro-
(Shi and Su, 2012).
mosome related function (Wood et al., 2008). Though a number of
studies have demonstrated the role of MCPH1 in HR, there exists a
lack of follow up studies to prove the role of MCPH1 in NHEJ. MCPH1 mediates G1/S and G2/M checkpoints

MCPH1 positively regulates the expression of BRCA1 and CHEK1

MCPH1 maintains telomeres
In mammalian cells, telomeres are protected by telosome com-
plexes (Blasco, 2003). A telosome complex constitutes proteins
such as TERF1 (telomere repeat factor-1), TERF2 (telomere repeat
factor-2), POT1 (protection of telomere-1), RAP1 (human analogue
of yeast telomeric protein rag1), TIN2 (TERF1 interacting protein)
and TPP1 (TINT1/PTOP/P1P1). Additionally, it is a platform for var-
ious proteins/signalling pathways to interact with telomeres (Xin
et al., 2008). Within the telosome complexes, TERF2 protein plays
an important role as it recruits many proteins, such as TINF2
(TERF1 [TRF1]-interacting nuclear factor 2), DCLRE1B (DNA cross-
link repair 1B) and PPP1R10 (protein phosphatase 1, regulatory
subunit 10), for the maintenance of the telomere ends (Kim et al.,
2009). In addition to recruitment of these proteins, TERF2 interacts
in U2OS cells (Xu et al., 2004; Lin et al., 2005; Passemard et al.,
2011)(Fig. 1). BRCA1 (breast cancer 1, early onset) and CHEK1 are
required for IR-induced S phase and G2/M cell cycle checkpoints
(Xu et al., 2001; Yarden et al., 2002). However, transcriptional
inactivation of BRCA1 and CHEK1 was not observed in human lym-
phoblastoid cell lines (LBLs) with different truncating mutations
in MCPH1 (MCPH1C74G and MCPH1427insA ) (Alderton et al., 2006).
These cells exhibited normal levels of phosphorylated CHEK1 and
an active ATR signalling, suggesting that clinical features of MCPH
syndrome patient cells cannot be attributed to MCPH1-dependent
transcriptional regulation of CHEK1 or BRCA1 (Alderton et al., 2006).
Knockdown of MCPH1 increased the radio-resistant DNA synthesis
(RDS) in U2OS cells (Xu et al., 2004). RDS is a phenomenon where
cells fail to inhibit DNA synthesis in response to IR (Rowley et al.,
1999). Moreover, depletion of MCPH1 lead to a higher content of
102 T. Venkatesh, P.S. Suresh / European Journal of Cell Biology 93 (2014) 98105
phospho-histone H3 (Ser 10), a mitotic marker in U2OS cells after
exposure to radiation (Xu et al., 2004). A higher staining of phospho-
histone H3 (Ser 10) indicates the presence of defective G2/M arrest.
Alderton et al. (2006) have studied the G2/M checkpoints in Lym-
phoblastoid cell lines (LBLs) from control (WT), an ATM-defective
ataxia telangiectasia (A-T), an ATR-Seckel and MCPH patients after
exposure to UV, and MCPH1 mutant cells were defective in G2/M
checkpoint. Also, MCPH1 depleted broblast cells derived from
patients show a delayed release from G2/M cell cycle checkpoints
following exposure to IR (Gavvovidis et al., 2010). Peripheral blood
cells of chronic myeloid leukemia (CML) patients show downreg-
ulated MCPH1 expression and exhibit defective G2/M arrest upon
exposure to hydroxyurea (HU) (Giallongo et al., 2011). Hence, the
effects of MCPH1 are different for IR and HU induced DNA damage
responses.
MCPH1 mediates apoptosis
It is well established that E2F1 upregulates genes such as
TP53 (tumor protein p53), TP73 (tumor protein p73), CASP3 (cas-
pase 3, apoptosis-related cysteine peptidase), CASP7 (caspase 7,
apoptosis-related cysteine peptidase) and APAF1 (apoptotic pepti-
dase activating factor 1) and hence, mediates apoptosis (Ginsberg,
2002). Knockdown of MCPH1 in HEK293 cells, results in decreased
expression of E2F1 target genes such as TP53, CASP3, CASP7, BRCA1
and CHEK1. Moreover, the second and the third BRCT domains
of MCPH1 interact with the N-terminal of E2F1 (Yang et al.,
2008). These observations indicate that MCPH1 is involved in E2F1
mediated apoptosis. Also, the ChIP and ChIP-re-ChIP assays have
conrmed the presence of E2F1-MCPH1 complex on the promoters
of CHEK1, BRCA1, TP73 and CASP7 in U2OS and HEK293 cells (Yang
et al., 2008). Interestingly, the E2F1 mediated regulatory effects of
human MCPH1 and rhesus macaque MCPH1 are different for p73,
Cyclin E and p14Arf (Shi et al., 2013). This indicates a functional
divergence of MCPH1 between human and non-human primates.
Effects of MCPH1 on mitotic phenotypes
The functions of MCPH1 have been an evolving area and a
new role for MCPH1 in the regulation of mitosis has been exam-
ined (Singh et al., 2012b). Singh et al. (2012b) have shown that
CDC27 interacts with MCPH1 using crystallization and ITC (isother-
mal titration calorimetry) based studies. CDC27 is a component of
anaphase promoting complex (APC). APC is very essential for the
regulation of mitosis and it is interesting to note that the knock-
down of CDC27 or MCPH1 exhibit similar mitotic phenotypes such
as presence of smaller nuclei, chromosomal segregation defects and
PCC (Singh et al., 2012b).
Insights into the functions of MCPH1 in cancer development
Cancer is a multifactorial disease caused by both genetic and
non-genetic factors. It is a major public concern and continues
to be the second common cause of death following heart dis-
eases in America (Valdivieso et al., 2012). Cancer is a phenomenon
Table 1
Summary of studies on MCPH1 in human cancer.
Status of MCPH1 Type of tumor
M, D, Me, LOH Breast
M Endometrial
D Primary ovarian cancer cultures, ovarian
D Glioblastoma
D, M, Me, LOH, miR Oral squamous cell carcinoma
Abbreviation: M, mutated; D, down regulated; Me, methylated; LOH; loss of heterozygosity; miR, miRNA regulated.
of dysregulation of gene expressions of oncogenes and tumor
suppressors. The dysregulation of gene expressions are either
a pre-transcriptional or post-transcriptional phenomenon. Pres-
ence of gene amplications/deletions, somatic mutation and DNA
hypo/hyper methylation are some of the mechanisms that result
in dysregulated gene expressions. Several DNA repair regulators
have been associated with human cancer such as BRCA1 and BRCA2
(Domchek et al., 2010). Hence, this opens up a novel perspective
that MCPH1 could have a functional role in cancer development or
its progression. But, there are no reports of signs of cancer predis-
position in the MCPH affected humans or knockout mice models. A
hint about the functional role for MCPH1 in cancer is provided by the
presence of 37 mutations in MCPH1 that has been recorded in COS-
MIC (Catalogue of somatic mutations in cancer) database (Forbes
et al., 2011; http://www.sanger.ac.uk/cosmic). Over a decadeof its
discovery, several studies have addressed the genetic, epigenetic
and functional roles of MCPH1 in various cancers. These studies
conducted over the past years have lead to a new and growing con-
cept that the story of MCPH1 does not end at primary microcephaly
and that it extends to the fascinating world of cancer too. Here, we
summarise the studies conducted to elucidate the role of MCPH1
in various human cancer.
MCPH1 in breast cancer
MCPH1 contains three BRCT domains, and it is well known that
breast cancer is inuenced by BRCT containing proteins like BRCA1
and BRCA2 (di Masi et al., 2011). Decreased copy number of MCPH1
has been reported in 38/54 breast cancer cell lines by CGH (com-
parative genomic hybridization) (Rai et al., 2006). MCPH1 is down
regulated in 70% (7/10) of breast cancer tissues by IHC (Rai et al.,
2008). Also, another IHC study has reported down regulation of
MCPH1 in 93/319 (29%) breast cancer samples (Richardson et al.,
2011) (Table 1). Reduced expression of MCPH1 has been demon-
strated in high grade, BRCA1 negative tumors and is a prognostic
indicator in breast cancer (Richardson et al., 2011). Also, presence
of T allele in the SNPs of MCPH1 (rs2912010 and rs1057090) is
associated with increasing tumor grade in breast cancer (Jo et al.,
2013). A 38 bp deletion in exon 10 of MCPH1 has been identied in
a breast cancer sample (Rai et al., 2006). Bhattacharya et al. (2013)
reported that reduced expression and alteration of ATM and MCPH1
were seen in 96% (121/ 126) of breast cancer samples and also,
patients with alterations in MCPH1 show poor outcome after treat-
ment with radiation or DNA interacting drugs (Bhattacharya et al.,
2013). Knockdown of MCPH1 lead to increased cellular prolifera-
tion and growth in soft agar in MCF10A (breast cancer) cells (Zhang
et al., 2013). In corroboration with these experiments, the ectopic
expression of MCPH1 in MCF7 (breast cancer) cells suppressed cell
proliferation and colony formation in vitro and tumor growth in
nude mice (Zhang et al., 2013). The mechanism of tumor suppres-
sion of MCPH1 is attributed to the increased stabilisation of the
master tumor suppressor TP53 (tumor protein p53) in MCF10A
cells (Zhang et al., 2013). MCPH1 induces auto-ubiquitination of
MDM2 (murine double minute 2), the negative regulator of TP53
References
Rai et al., 2006; Richardson et al., 2011; Bhattacharya et al., 2012.
Bilbao et al., 2010
Rai et al., 2006; Brning-Richardson et al., 2011
Hagemann et al., 2008
Venkatesh et al., 2013
 T. Venkatesh, P.S. Suresh / European Journal of Cell Biology 93 (2014) 98105
which eventually leads to the proteosomal degradation of MDM2
and thus, stabilizes TP53 (Zhang et al., 2013).
MCPH1 in endometrial cancer
Endometrial cancer is the most common malignancy of the
female genital tract (Siegel et al., 2012). It is estrogen dependent and
patients with advanced stage cancers have a much less favorable
prognosis (Karaayvaz et al., 2012). Mononucleotide deletions in the
A tracts of exons 4 (2/41 samples), 5 (1/41 samples) and 8 (2/41
samples) of the MCPH1 gene have been reported in microsatellite
instability (MSI) positive endometrial cancer tissues, accounting to
a total mutation rate of 12% (5/41) (Bilbao et al., 2010). All these
mutations were monoallelic and 62 microsatellite stable (MSS)
endometrial cancer tissues were normal for MCPH1 (Bilbao et al.,
2010).
MCPH1 in ovarian cancer
Ovarian cancer is the fth-leading cause of cancer-related
death in the United States, and is the most lethal of all gyne-
cological malignancies (Zhan et al., 2013). Rai et al. (2006) have
shown decreased MCPH1 genomic DNA copy number in 35/87
(40.22%) advanced epithelial ovarian cancers using CGH (Compar-
ative genomic hybridization). Also, decreased MCPH1 level was
seen in 19/30 ovarian cancer specimens in comparison to seven
benign ovarian specimens (Rai et al., 2006)(Table 1). Additionally,
ICC (immunocytochemistry) of 36 primary cultures from ovarian
cancer samples showed a strong/moderate staining of MCPH1 in
the nucleus and out of these, a weak cytoplasmic staining was seen
in 25% of the samples (Brning-Richardson et al., 2011).
MCPH1 in glioblastoma
Glioblastoma multiforme (GBM) is the most prevalent malig-
nant brain tumor in adults. These tumors may occur as de novo
or progress from a low grade astrocytoma (LGA, WHO grade II)
or anaplastic astrocytoma (WHO grade III) (Biernat et al., 1997).
Hagemann et al. (2008) analysed the transcript level of MCPH1 in
15 LGA and 15 GBM tumors. They reported that all the normal brain
samples expressed MCPH1, but all at low levels. MCPH1 expression
was reduced to 20% and 27% in LGA and GBM tumors, respec-
tively, when compared to the normal brain tissues (Hagemann et al.,
2008).
MCPH1 in oral squamous cell carcinoma
Oral squamous cell carcinoma (OSCC) is a subgroup of HNSCC
(Supic et al., 2009). It accounts for 24% of all malignancies in the
west. However, in Indian subcontinent, OSCC accounts to 4050%
of all malignancies (Mehrotra et al., 2006). Microsatellite analy-
sis has shown LOH (loss of heterozygosity) at the D8S518 and
D8S277 markers anking the MCPH1 locus in 1/21 oral tumors
(Wu et al., 1997). Venkatesh et al. (2013) have shown LOH
of MCPH1 in 14/71 (19.72%) informative samples. Also, a pro-
tein truncating mutation c.1561G.T(p.Glu521X) was identied in
1/15 OSCC samples (Venkatesh et al., 2013). Furthermore, their
study has reported that MCPH1 was down regulated at both
the transcript and protein levels in 21/41 (51.22%) and 19/25
(76%) OSCC samples, respectively (Table 1). Overexpression of
MCPH1 in KB cells decreased cellular proliferation, anchorage-
independent growth in soft agar, cell invasion and tumor size in
nude mice, indicating its tumor suppressive function. Additionally,
it is intriguing to note that MCPH1 is regulated by miR-27a in OSCC
(Venkatesh et al., 2013).
103
Implication of MCPH1 in Otitis media (OM)
Recently, a novel role for MCPH1 in the development of otitis
media has been identied. Otitis media is a multifactorial disease
characterized by the inammation of the middle ear. Environmen-
tal factors such as smoking and type of child care, anatomical
dysmorphology, Eustachian-tube function, viral and bacterial load,
and, genetic predisposition are some of the factors responsible
for the development and persistence of OM (Chen et al., 2013).
Chen et al. (2013) generated a Mcph1 decient mice and examined
the hearing ability in these mice using ABR (auditory brainstem
response) analysis. Homozygous mutant mice exhibited elevated
ABR thresholds as early as 3 weeks of age. The heterozygous mice
showed no hearing impairment in consistent with the recessive
model of inheritance for patients with MCPH1 mutations (Chen
et al., 2013). Furthermore, anatomical and histological studies in
these Mcph1 decient mice revealed OM. Several defects in the mid-
dle ear such as thickened and white bone forming the bulla instead
of the normal thin and transparent bone, retracted tympanic mem-
brane, bubbles present underneath the tympanic membrane, or
middle ear cavities lled with clear or cloudy uid were detected
(Chen et al., 2013). Chen et al. (2013) performed immunohisto-
chemical studies and revealed that Mcph1 is expressed in the
middle ear of the wildtype mice. Still, question remains regarding
the MCPH1 signalling pathways that lead to OM.
Conclusions
Studies so far have proved that MCPH1 has roles in the develop-
ment of brain, cancer and OM. Though several interacting proteins
and pathways have been elucidated for MCPH1, there are sev-
eral interesting aspects of MCPH1 which are yet to be unravelled.
They include elucidation of a more direct centrosomal function for
MCPH1 or to identify whether MCPH1 interacts with any other
MCPH proteins. Our growing familiarity with MCPH proteins has
proved their signalling to be highly complex in the pathogenesis of
MCPH. However, a possibility of the existence of a master regula-
tor for all these MCPH proteins cannot be ruled out. Also, the area
less visited is the post translation modication of MCPH1, and how
these could inuence the functions of MCPH1 in primary micro-
cephaly, cancer and OM. Wide scale systems biology, knock out
studies and networking based approaches will enhance our knowl-
edge of using MCPH1 as a therapeutic target for cancer. As, MCPH1
is a negative regulator of MDM2 in breast cancer, molecular effec-
tors that can up regulate MCPH1 can be an attractive anticancer
therapeutic approach.
Conict of interest
The authors declare no conict of interest.
Acknowledgement
We would like to thank Professor Arun Kumar, Indian Institute
of Science, Bangalore for his help and support and also the study
institutes. We thank Kavita Bommasamudram, Stony Brook Uni-
versity, New York for critical reading of the manuscript. We also
thank the reviewers for their valuable suggestions to improve the
manuscript.
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