Evolution of primary microcephaly genes and the enlargement
of primate brains
Chris Ponting1 and Andrew P Jackson2
Brain size, in relation to body size, has varied markedly during
the evolution of mammals. In particular, a large cerebral cortex
is a feature that distinguishes humans from our fellow primates.
Such anatomical changes must have a basis in genetic
alterations, but the molecular processes involved have yet to be
defined. However, recent advances from the cloning of two
human disease genes promise to make inroads in this
important area. Microcephalin (MCPH1) and Abnormal spindle-
like microcephaly associated (ASPM) are genes mutated in
primary microcephaly, a human neurodevelopmental disorder.
In this atavistic condition, brain size is reduced in volume to a
size comparable with that of early hominids. Hence, it has been
proposed that these genes evolved adaptively with increasing
primate brain size. Subsequent studies have lent weight to this
hypothesis by showing that both genes have undergone
positive selection during great ape evolution. Further functional
characterisation of their proteins will contribute to an
understanding of the molecular and evolutionary processes
that have determined human brain size.
Addresses
1
MRC Functional Genetics, University of Oxford, Department of Human
Anatomy and Genetics, South Parks Road, Oxford OX1 3QX, UK
2
MRC Human Genetics Unit, Western General Hospital, Crewe Road,
Edinburgh, EH4 2XU, UK
Corresponding authors: Ponting, Chris (chris.ponting@anat.ox.ac.uk);
Jackson, Andrew P (andrew.jackson@hgu.mrc.ac.uk)
Current Opinion in Genetics & Development 2005, 15:241248
This review comes from a themed issue on
Genetics of disease
Edited by Veronica van Heyningen and David FitzPatrick
Available online 22nd April 2005
0959-437X/$ see front matter
# 2005 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.gde.2005.04.009
Introduction
There has been a dramatic expansion in brain size during
mammalian evolution, with a thousand fold increase in
cortical surface area between mouse and man [1]. Morpho-
metric studies (see Glossary) have revealed a scaling of the
brain relative to body size during mammalian evolution
[2], with disproportionate enlargement of the cerebral
cortex [3]. In primates, this might have been driven by
increased social, rather than environmental, complexity
[4]. Expansion has been through increased surface area
rather than increased cortical thickness [1,2]. This has
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been accommodated topologically by folding of the cere-
bral cortex to form convolutions, so that there is a transition
from the smooth cortical surface seen in rodents to one of
multiple folds (gyri and sulci; see Glossary) in humans.
This review contrasts the pathological reduction in
human brain size (primary microcephaly) with the
increase in relative brain size during primate evolution.
We describe the primary microcephaly genes and their
functions, and address the recently proposed roles of
ASPM and microcephalin genes in determining brain
size among different mammals.
Cellular determinants of brain size
The relative rates of proliferation and cell death occurring
during neurogenesis (see Glossary) are likely to be major
determinants of mammalian brain size. Evolutionary
expansion of the mammalian brain must, at least in part,
be the result of increased numbers of cell divisions during
neurogenesis [5]. An evolutionary increase in the number
of columnar functional units generated by proliferative
units of progenitor cells has been proposed to explain
how increased cell number translates into expansion in
cortical surface area [6].
Neurogenesis first generates a pool of progenitor cells by
symmetric cell division, and neurons are subsequently
formed by asymmetric cell divisions. It has been proposed
that small changes in the relative proportion of such
divisions could be responsible for marked alterations in
cortical surface area [7]. Indeed, it has been demonstrated
recently that increasing the proportion of cycling progeni-
tor cells is sufficient to increase cortical surface area,
producing a mouse cerebral cortex with sulci and gyri
[8]. Widespread neuronal cell death also occurs during
foetal brain development, and the amount of cell death is
also an important determinant of brain size. Inactivating
components of the apoptotic pathway also leads to an
enlarged sulcated (i.e. furrowed) mouse brain [9,10].
Candidate genes
Despite insights into the cellular mechanisms that regulate
brain size, the molecules and genes responsible for this
process remain enigmatic. In general, genes expressed
predominantly in the brain have evolved, between human
and mouse, more slowly than those expressed in other
tissues [11]. Nevertheless, some nervous system genes
exhibit more rapid evolution among primates than among
rodents [12], and so changes in these genes might have
been responsible for brain size modifications. Clearly,
molecules involved in regulating cellular proliferation or
Current Opinion in Genetics & Development 2005, 15:241248
242 Genetics of disease
Glossary
Aneuploidy Condition of having an abnormal number of
chromosomes.
Atavistic Regarding a reversion to an ancestral form.
Early truncating mutations DNA mutations that give rise to a stop
codon in the 50 -most region of a protein coding sequence.
Gyri Elevations in the surface of the cerebral hemispheres.
IQ domain A protein motif often containing conserved isoleucine
and glutamine amino acids.
Morphometric Regarding the measurement of shape and size.
Neurogenesis The developmental process of producing neural
cells.
Sulci Furrows in the surface of the cerebral hemisphere.
apoptosis are likely candidates, given the processes known
to influence brain size outlined above.
It has been suggested that genes responsible for human
neurogenetic disorders might be of particular interest in
this regard because, through disease, they have demon-
strated the potential to alter developmental outcome of
brain morphology and function [13

]. In respect to brain
size, genes for primary microcephaly are prime candidates.
Primary microcephaly an evolutionary
condition?
Primary microcephaly is a Mendelian neurodevelopmen-
tal disorder in which there is marked reduction in human
brain size. Brain volume is reduced to a third of normal
[14], to a size comparable with that of early hominids
(Figure 1) [15]. Surprisingly, in this disorder, the great loss
in volume of cerebral cortex does not lead to significant
neurological dysfunction; apart from a moderate reduc-
tion in cognitive ability [16]. The cerebral cortex has a
simplified gyral pattern [17

,18], with no major abnorm-
ality of cortical architecture [14,19]. These findings, along
with the facial characteristics of a hypoplastic skull vault
and sloping forehead, have led to the proposal that
primary microcephaly might be an evolutionary (i.e.
atavistic) condition (see Glossary) [19,20].
Microcephaly genes
Linkage mapping studies in consanguineous families
have identified six gene loci for primary microcephaly
(MCPH16) [2127]. Subsequently, two of these genes
were positionally cloned: first microcephalin [28] and then
abnormal spindle-like microcephaly-associated (ASPM) [18],
for the MCPH1 and MCPH5 loci, respectively. Mutations
in ASPM appear to be a major cause of primary micro-
cephaly [2931], whereas microcephalin mutations have
only been reported in three families [17

,28].
Microcephalin
Microcephalin (MCPH1) is an 835 amino acid protein
containing BRCA1 C-terminal (BRCT) domains [28]. Its
closest human homologues are topoisomerase II-binding
protein (TopBP1) and BRCA1. Three BRCT domains
are predicted to be present [32], one at the N-terminus
Current Opinion in Genetics & Development 2005, 15:241248
and a further pair at the C-terminus (Figure 2a). BRCT
domains occur in many DNA repair and cell cycle pro-
teins, and they participate in proteinprotein and pro-
teinDNA interactions [33]. The functions of these
domains have recently been further clarified by the
demonstration that they specifically bind proteins phos-
phorylated on serine and/or threonine residues [34]. The
implication is, thus, that their binding is modulated by
phosphorylation as a result of either cell cycle stage-
specific events or signals arising from DNA damage. This
year, a role in DNA damage response was confirmed for
microcephalin, after the demonstration that microcepha-
lin localises to DNA repair foci and that cells depleted for
microcephalin by RNA interference exhibit failure in cell
cycle checkpoints [35

]. This gene has also been isolated
(as BRIT1) in a screen for negative regulators of telomer-
ase [36
], and has a demonstrated role in the cell cycle in
regulating chromosome condensation [17

].
Both mutations identified in microcephalin have been early
truncating mutations (see Glossary) and, therefore, are
predicted to lead to a complete loss of gene function [28].
Given the diverse roles outlined above, it remains unclear
how abrogation of microcephalin leads to reduction in
brain size. Loss of DNA repair molecules can lead to
increased cell death during neurogenesis [37]. Conver-
sely, embryonic proliferation might be reduced, as in the
case of Brca1/ mice [38].
ASPM
As with microcephalin, ASPM was positionally cloned
using consanguineous Pakistani families, and locus refine-
ment by identifying shared ancestral haplotypes [18]. All
21 mutations reported to date have been protein-truncat-
ing, and are distributed throughout the gene [30,31],
presumably implying that primary microcephaly is the
consequence of functionally null mutations.
ASPM is a very large protein of 3477 amino acids, and
comprises a putative microtubule-binding domain at its N-
terminal, a calponin-homology domain, multiple IQ cal-
modulin-binding domains (see Glossary) and a terminal
region (Figure 2b). With the exception of some rodents, the
number of ASPM IQ domains is constant among diverse
mammals [39
]. This contradicts an early proposal [18] that
IQ domain number positively correlates with increased
nervous system complexity. Like microcephalin, ASPM is
expressed in the cerebral cortical ventricular zone and
proliferative zones of the medial and lateral ganglionic
eminences during neurogenesis [18,28]. ASPM is also
expressed postnatally in regions of persistent neurogenesis.
Although the cellular functions of ASPM have yet to be
defined, its fly orthologue, abnormal-spindle protein
(Asp), has been extensively studied and has roles in both
mitosis and meiosis [4043]. Asp is necessary for the
centrosome to function as a microtubule-organising
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 Evolution of primary microcephaly genes and the enlargement of primate brains Ponting and Jackson 243

Figure 1

Evolutionary expansion of the primate brain contrasts with the pathological reduction in brain size seen in primary microcephaly. (a) Primate skulls
from the Museum of Comparative Zoology, Harvard University (Reproduced with permission from [54]).There has been a large increase in brain
size during primate evolution, reflected by enlargement of the skull vault and cranial capacity. (b) Magnetic resonance imaging of a normal adult
human head (left) and an individual with MCPH1 primary microcephaly (right). Midline sagittal (T1 weighted) sections; adapted with permission
from [28]). In primary microcephaly, there has been a marked decrease in brain volume. In particular, the cerebral cortex is much smaller and
exhibits reduced cortical folding with a simplified gyral pattern.

centre and to focus the poles of the mitotic spindle [40]. It
is also required for cytokinesis, where it is an organiser of
the central spindle [41]. Mutation of asp leads to mitotic
arrest in larval neuroblasts [42].
Positioning of the mitotic spindle perpendicular to the
epithelial surface is a regulated process in fly neuroblasts
and is required for correct orientation of cell cleavage
during asymmetric cell division [44]. Additionally, clea-
vage plane orientation might be important in localising
asymmetric determinants in mammals [45]. Given the
known functions of Asp, it was, therefore, proposed that
mutation of ASPM might reduce brain size by influencing
mitotic spindle orientation, and reduce neural cell num-
ber through an effect on the ratio of asymmetric to
symmetric cell divisions [18].
MCPH1 is ubiquitous in metazoans
A previous report [13

] suggested that MCPH1 is found
only in mammals. By contrast, we believe that MCPH1
orthologues are likely to be present in all chordates,
because these genes are identifiable in all primates,
carnivores, ruminants, marsupials, birds, fish, amphibians
and sea squirts for which we currently possess genomic
sequence. These MCPH1 sequences are each identical in
architecture to human MCPH1. Their N-terminal BRCT
domain is separated from two C-terminal BRCT domains
by a long stretch of inter-BRCT domain sequence (IBS)
that appears devoid of known domain structures and that
is mostly encoded by a single exon.
Similar MCPH1 genes are also present in other metazoan
species, in particular the two main invertebrate model
organisms, Drosophila melanogaster (gene CG8981) and
Caenorhabditis elegans (W04A8.1).
The MCPH1 gene, however, might have arisen in eukar-
yotic evolution even earlier than the earliest metazoan.
Other transcripts are known, from more distantly-related
eukaryotic species, that all encode sequences similar to

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244 Genetics of disease

Figure 2

(a) Microcephalin

Exons
1 2 3 4 5 6 7 8 9 10 11 12 13 14

Domains

BRCT IBS BRCT BRCT

(b) ASPM

Exons

1 2 3 13 17 18 28

Domains

Microtubule binding Calponin IQ (multiple) C terminus

homology

Current Opinion in Genetics & Development

Exon structure of microcephalin and ASPM genes (a) Microcephalin is an 835 amino acid (aa) protein, mutated in MCPH1 primary microcephaly.
The protein is predicted to contain three BRCA1 C-terminal (BRCT) domains. An N-terminal BRCT domain is separated by over 500 aa of
inter-BRCT domain sequence (IBS), which has no predicted domain structure, from a pair of BRCT domains at the C-terminus. Regions of the
gene have undergone high levels of adaptive evolution (red exons, KA/KS > 3, among ancestors of old world monkeys and great apes), whereas
purifying selection has occurred particularly in the gene regions encoding BRCT domains. Exon structure and KA/KS ratios are taken from [13

],
and predicted domain structure from [28].(b) Abnormal spindle-like microcephaly-associated (ASPM) protein is a very large molecule of 3477
aa divided into four regions: a N-terminal putative microtubule binding domain (purple), calponin-homology domain (orange), multiple IQ,
calmodulin-binding, domains (red) and a C-terminal region (blue) [18]. Exons 3 and 18 have undergone adaptive evolution, with regions of
significantly elevated KA/KS in the ape lineages leading to humans (red exons) [39
].

the BRCT domain-containing regions of human MCPH1.
These include a fungus (Cryptococcus neoformans; gene
CNBK1030), an amoeba (Entamoeba histolytica), and a
plant (Sorghum bicolor; EST [expressed sequence tag]
CD430938). However, full-length MCPH1-like sequences
containing N- and C-terminal BRCT domains, separated
by an IBS region, have yet to be identified outside of the
metazoans.
Adaptive evolution of microcephalin
When human and mouse sequences are aligned, MCPH1
appears among the top 10% of rapidly evolving genes:
they have accumulated more amino acid-changing (non-
synonymous) nucleotide substitutions for each silent
(synonymous) substitution than most other genes. Such
rapid evolution can occur neutrally that is, as a result of
reduced functional constraint on sequence or by posi-
tive selection, caused by the accumulation of nonsynon-
ymous changes that confer adaptive benefit.
Recently, two studies have attempted to distinguish posi-
tive selection from neutrality during the evolution of
MCPH1 genes [13

,46
]. They demonstrated that
whereas the flanking BRCT domains have accumulated
amino acid changes relatively slowly, as befits sequence
that is well-conserved back to at least the earliest metazoan
organism, the intervening IBS has acquired substantial
numbers of amino acid alterations. This is also consistent
with the considerably reduced sequence similarity
between, for example, sea squirt (Ciona intestinalis) and
mammalian MCPH1 genes in their extended IBS regions,
or even between different Drosophila species (see below).

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 Evolution of primary microcephaly genes and the enlargement of primate brains Ponting and Jackson 245

Using sequence differences in MCPH1 genes, both within
the human population and among different mammals,
these investigators demonstrated that although neutral
evolution could explain some of their changes Darwinian
positive selection most likely also contributed changes.
These occurred specifically along the simian lineages that
led to modern apes and Old World monkeys. Similar
conclusions were drawn from human polymorphism data
[13

,46
].
By contrast, the evolution of MCPH1 genes in species
other than primates appears to have been more pedes-
trian, because there is no statistical evidence for Darwi-
nian positive selection for any of these other animals.
Figure 3
Nevertheless, the extended IBS of cat and fly MCPH1
genes have each independently acquired different
repeats (Figure 3). In most Drosophila species, there
are three such repeats, but Drosophila pseudoobscura has
acquired an extra pair, making five in total. These repeats
have been acquired relatively recently, certainly in the 55
million years since the last common ancestor of cat with
dog [47] and the 25 million years since the last common
ancestor of D. pseudoobscura and the other Drosophila
species [48]. Therefore, it is possible that these repeats
were acquired adaptively by flies and cats. The fruit fly
Mcph1 protein repeats are distinguished from those of the
mosquito protein by their different conservation of serine
and threonine residues, respectively (Figure 3).

(a)
RRR*
D. melanogaster 1: PVRTTRRRSCVAEI 436-449
D. melanogaster 2: DTRMTRRRSSLLTS 456-469
D. melanogaster 3: EPRMTRRRSSLLGI 484-497
D. pseudoobscura 1: PVRTSRRRSSAHIS 437-450
D. pseudoobscura 2: EQRVTRRRSSAHIP 464-477
D. pseudoobscura 3: EPRMTRRRSSAHIL 515-528
D. pseudoobscura 4: EPRLKRRRSSAHIP 542-555
D. pseudoobscura 5: DLRMTRRRSSLLQA 569-582
D. yakuba 1: PVRTTRRRSCVAEI 439-452
D. yakuba 2: DTRMTRRRSSLLTS 459-472
D. yakuba 3: EPRMTRRRSSLLAT 487-500
D. ananassae 1: PVRTNRRRSCAAEI 437-450
D. ananassae 2: EPRMTRRRSSLHIN 454-467
D. ananassae 3: EPRMTRRRSSLHLA 481-494
D. mojavensis 1: PLRQLRRRSLAYVC 302-315
D. mojavensis 2: GSRLTRRRSQLNIT 328-341
D. mojavensis 3: EPRITRRRSLQLLT 364-378
D. virilis 1: PLRASRRRSSACGA 311-324
D. virilis 2: EPRMTRRRSSLLSM 337-350
D. virilis 3: EPRMPRRRSSLQLA 372-385
A. gambiae 1: GTPRGRRKTVMFTP 459-472
A. gambiae 2: PKQTFRRKTIVLPN 495-508
A. gambiae 3: SNTSNRRKTIVAAP 520-533
A. gambiae 4: VRVSSRRKTIVAAK 601-614
A. gambiae 5: KNKRSRRKTIAFGN 632-645
A. gambiae 6: NAPKDRRRTLFIPT 726-739
A. gambiae 7: LSLINRRRTLYTPS 762-775
A. gambiae 8: RAPDRRRRTVFDIT 858-871

(b)

Felis catus 1: LKTGSVHSSAPAGPSAS 274-290

Felis catus 2: LKTGSVHSSAPAGPSAS 291-307
Felis catus 3: LKTGSVHSLAPAGHLSQ 308-324
Canis familiaris LKTSCVPSLAPPGYLCQ 275-291

Current Opinion in Genetics & Development

Amino acid conservation of MCPH1 proteins (a) Repetitive arginine- (R) and serine- (S) or threonine- (T) rich motifs in arthropod MCPH1 proteins.
Note that Drosophila pseudoobscura MCPH1 possesses five repeats, whereas the other drosophilids possess three, and the mosquito Anopheles
gambiae MCPH1 possesses eight. In addition, Drosophila repeats conserve a serine in the position that the mosquito repeats always contain a
threonine. Alignment columns shaded in light grey contain at least 80% hydrophobic amino acids. (b) Triplicated repeat in cat (Felis catus)
MCPH1 that is present as a single copy in all other mammals known including dog (Canis familiaris).

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246 Genetics of disease
Given this pattern of conservation, and the fact that
BRCT domains are phosphoserine- or phosphothreo-
nine-binding domains [34,49], we speculate that fly,
and possibly human, microcephalin function could be
regulated by phosphorylated serines or threonines in
the IBS repeats. Such residues might well interact with
the BRCT domains of other proteins, or even interact
with the flanking N- or C-terminal BRCT domains of
microcephalin itself.
Adaptive evolution of ASPM genes
The ASPM gene also appears to have accumulated sig-
nificant numbers of adaptive changes in the terminal
human evolutionary branch since the last common ances-
tor of chimpanzees and humans [39
,50
]. By comparing
ASPM gene sequences, gorillas, chimpanzees and humans
were distinguished from other primates by having accu-
mulated large numbers of nonsynonymous nucleotide
substitutions in this gene [39
,50
]. Moreover, during
the human lineage since the last common ancestor with
chimpanzees, approximately 15 of these substitutions in
this gene have been adaptive, and these are proposed to
have contributed to the progressive enlargement of the
hominid cerebral cortex [39
]. Significantly more amino
acid changes have occurred between chimpanzee and
human sequences than they have recently within the
human population, which again argues for positive selec-
tion in the terminal human branch [39
,50
].
Among apes, the largest concentrations of nonsynon-
ymous substitutions occur within two long exons (3
and 18) that encode large portions of likely microtu-
bule-binding and calmodulin-binding regions, respec-
tively [39
,51

] (Figure 2). The ASPM calponin-
homology domain has experienced substantially fewer
such substitutions. Hence, for both ASPM and MCPH1
gene sequences, it is the extended exons that have
accumulated disproportionately large numbers of nonsy-
nonymous substitutions, whereas regions encoding large
folded protein domains have attracted relatively few. It is
also noteworthy that adaptive sites are also clustered in a
large exon (exon 11) of BRCA1 [52] that provides much of
the sequence intervening between its terminal structural
domains. We believe it improbable that large exons are
more likely, in general, to accumulate adaptive mutations.
Rather, such exons (exon 8 of microcephalin and exon 18 of
ASPM) might have been extended by successive internal
duplications over a time period that was sufficiently short
to have not suffered from intron insertion.
How might microcephaly genes have been
involved in evolution of human brain size?
Pathological changes in microcephalin and ASPM reduce
brain size, therefore demonstrating that these genes have
the capability of altering the developmental outcome of
neurogenesis. These mutations apparently recapitulate
an earlier hominid phenotypic state. It is clear, however,
Current Opinion in Genetics & Development 2005, 15:241248
that these modern pathological changes, which are loss-
of-function mutations, do not recapitulate the ancient
hominid genotype. Instead microcephalin and ASPM genes
have been preserved as functional sequences, without
degenerating into pseudogenes since at least an early
metazoan ancestor to extant animals.
In general, microcephalin or ASPM gene function does not
promote enlargement of the neocortex, because this
structure is specific to mammals, and the origin of each
of these genes predates the first mammal. Instead, incre-
mental adaptive changes to these genes might have subtly
altered cell cycle proliferation or reduced the propensity
of neurons to apoptosis. For ASPM, alterations in mitotic
spindle function, perhaps as a consequence of altered
spindle orientation, might have increased symmetric cell
division at the expense of asymmetric cell division in
neurogenesis, thereby increasing the eventual neuronal
number. Alternatively, it is suggested that sequence
changes might have resulted in increased neuronal survi-
val, through more robust chromosome segregation [51

]
resulting from reduction in the high levels of aneuploidy
(see Glossary) seen in rodent neuroblasts [53].
Conclusions
Despite answers to the question of whether primate
microcephalin or ASPM genes evolved adaptively, it still
remains to be established why they did so. The expres-
sion of these genes is not restricted to the central nervous
system and, because their known functions are not con-
fined to processes that regulate brain morphology, it is still
possible that their evolution is causally unconnected with
the expansion of the cerebral cortex in great apes. Clearly,
an improved understanding of the function of these
proteins would also assist in resolving this issue, as would
analyses of the evolutionary histories of other primary
microcephaly genes, once they have been identified. We
wait with interest for developmental and cellular studies
that will address the biological significance of the bio-
chemical changes identified from these evolutionary stu-
dies.
Acknowledgements
This work was funded by the UK Medical Research Council. We thank
Ian Hope for bringing the C.elegans MCPH1 orthologue to our attention.
References and recommended reading
Papers of particular interest, published within the annual period of
review, have been highlighted as:

of special interest


of outstanding interest
1. Rakic P: A small step for the cell, a giant leap for mankind:
a hypothesis of neocortical expansion during evolution.
Trends Neurosci 1995, 18:383-388.
2. Northcutt RG, Kaas JH: The emergence and evolution of the
mammalian neocortex. Trends Neurosci 1995, 18:373-379.
3. Clark DA, Mitra PP, Wang SS: Scalable architecture in
mammalian brains. Nature 2001, 411:189-193.
www.sciencedirect.com
 Evolution of primary microcephaly genes and the enlargement of primate brains Ponting and Jackson 247
4. Dunbar RIM: Coevolution of neocortical size, group size and
language in humans. Behav Brain Sci 1993, 16:681-735.
5. Kornack DR, Rakic P: Changes in cell-cycle kinetics during the
development and evolution of primate neocortex. Proc Natl
Acad Sci USA 1998, 95:1242-1246.
6. Rakic P: Specification of cerebral cortical areas. Science 1988,
241:170-176.
7. Caviness VS Jr, Takahashi T, Nowakowski RS: Numbers, time
and neocortical neuronogenesis: a general developmental
and evolutionary model. Trends Neurosci 1995, 18:379-383.
8. Chenn A, Walsh CA: Regulation of cerebral cortical size by
control of cell cycle exit in neural precursors. Science 2002,
297:365-369.
9. Kuida K, Zheng TS, Na S, Kuan C, Yang D, Karasuyama H,
Rakic P, Flavell RA: Decreased apoptosis in the brain and
premature lethality in CPP32- deficient mice. Nature 1996,
384:368-372.
10. Kuida K, Haydar TF, Kuan CY, Gu Y, Taya C, Karasuyama H,
Su MS, Rakic P, Flavell RA: Reduced apoptosis and cytochrome
c-mediated caspase activation in mice lacking caspase 9.
Cell 1998, 94:325-337.
11. Winter EE, Goodstadt L, Ponting CP: Elevated rates of protein
secretion, evolution, and disease among tissue-specific
genes. Genome Res 2004, 14:54-61.
12. Dorus S, Vallender EJ, Evans PD, Anderson JR, Gilbert SL,
Mahowald M, Wyckoff GJ, Malcom CM, Lahn BT: Accelerated
evolution of nervous system genes in the origin of Homo
sapiens. Cell 2004, 119:1027-1040.
13. Evans PD, Anderson JR, Vallender EJ, Choi SS, Lahn BT:


Reconstructing the evolutionary history of microcephalin,
a gene controlling human brain size. Hum Mol Genet 2004,
13:1139-1145.
These investigators showed that the microcephalin amino acid sequence
has been modified adaptively during its evolution from simian ancestors
to humans and chimpanzees.
14. Mcreary BD, Rossiter JP, Robertson DM: Recessive (true)
microcephaly: a case report with neuropathological
observations. J Intellect Disabil Res 1996, 40:66-70.
15. Wood B, Collard M: The human genus. Science 1999,
284:65-71.
16. Bundey S: Microcephaly. In Genetics and Neurology, 2nd edn.
Edited by: Churchill Livingstone; 1992:20-24. Emery AEH (Series
Editor): Genetics in Medicine and Surgery.
17. Trimborn M, Bell SM, Felix C, Rashid Y, Jafri H, Griffiths PD,


Neumann LM, Krebs A, Reis A, Sperling K et al.: Mutations in
microcephalin cause aberrant regulation of chromosome
condensation. Am J Hum Genet 2004, 75:261-266.
Patients with microcephaly caused by mutations in microcephalin were
shown in this study to have an associated cellular phenotype of aberrant
regulation in chromosome condensation. Cells deficient in microcephalin
spend extended periods of time with chromosome condensation in the G2
phase of the cell cycle. Furthermore, chromosome condensation con-
tinues for some time post-mitosis.
18. Bond J, Roberts E, Mochida GH, Hampshire DJ, Scott S,
Askham JM, Springell K, Mahadevan M, Crow YJ, Markham AF
et al.: ASPM is a major determinant of cerebral cortical size.
Nat Genet 2002, 32:316-320.
19. Mochida GH, Walsh CA: Molecular genetics of human
microcephaly. Curr Opin Neurol 2001, 14:151-156.
20. Komai T, Kishimoto K, Ozaki Y: Genetic study of microcephaly
based on Japanese material. Am J Hum Genet 1955, 7:51-65.
21. Jackson AP, McHale DP, Campbell DA, Jafri H, Rashid Y,
Mannan J, Karbani G, Corry P, Levene MI, Mueller RF et al.:
Primary autosomal recessive microcephaly (MCPH1) maps
to chromosome 8p22-pter. Am J Hum Genet 1998,
63:541-546.
22. Jamieson CR, Govaerts C, Abramowicz MJ: Primary autosomal
recessive microcephaly: homozygosity mapping of MCPH4 to
chromosome 15. Am J Hum Genet 1999, 65:1465-1469.
www.sciencedirect.com
23. Jamieson CR, Fryns JP, Jacobs J, Matthijs G, Abramowicz MJ:
Primary autosomal recessive microcephaly: MCPH5 maps to
1q25-q32. Am J Hum Genet 2000, 67:1575-1577.
24. Leal GF, Roberts E, Silva EO, Costa SM, Hampshire DJ,
Woods CG: A novel locus for autosomal recessive primary
microcephaly (MCPH6) maps to 13q12.2. J Med Genet 2003,
40:540-542.
25. Moynihan L, Jackson AP, Roberts E, Karbani G, Lewis I, Corry P,
Turner G, Mueller RF, Lench NJ, Woods CG: A third novel locus
for primary autosomal recessive microcephaly maps to
chromosome 9q34. Am J Hum Genet 2000, 66:724-727.
26. Pattison L, Crow YJ, Deeble VJ, Jackson AP, Jafri H, Rashid Y,
Roberts E, Woods CG: A fifth locus for primary autosomal
recessive microcephaly maps to chromosome 1q31. Am J Hum
Genet 2000, 67:1578-1580.
27. Roberts E, Jackson AP, Carradice AC, Deeble VJ, Mannan J,
Rashid Y, Jafri H, McHale DP, Markham AF, Lench NJ et al.: The
second locus for autosomal recessive primary microcephaly
(MCPH2) maps to chromosome 19q13.1-13.2. Eur J Hum Genet
1999, 7:815-820.
28. Jackson AP, Eastwood H, Bell SM, Adu J, Toomes C, Carr IM,
Roberts E, Hampshire DJ, Crow YJ, Mighell AJ et al.:
Identification of microcephalin, a protein implicated in
determining the size of the human brain. Am J Hum Genet 2002,
71:136-142.
29. Roberts E, Hampshire DJ, Pattison L, Springell K, Jafri H, Corry P,
Mannon J, Rashid Y, Crow Y, Bond J et al.: Autosomal recessive
primary microcephaly: an analysis of locus heterogeneity and
phenotypic variation. J Med Genet 2002, 39:718-721.
30. Bond J, Scott S, Hampshire DJ, Springell K, Corry P,
Abramowicz MJ, Mochida GH, Hennekam RC, Maher ER, Fryns JP
et al.: Protein-truncating mutations in ASPM cause variable
reduction in brain size. Am J Hum Genet 2003, 73:1170-1177.
31. Kumar A, Blanton S, Babu M, Markandaya M, Girimaji S: Genetic
analysis of primary microcephaly in Indian families: novel
ASPM mutations. Clin Genet 2004, 66:341-348.
32. Schultz J, Milpetz F, Bork P, Ponting CP: SMART, a simple
modular architecture research tool: identification of signaling
domains. Proc Natl Acad Sci USA 1998, 95:5857-5864.
33. Huyton T, Bates PA, Zhang X, Sternberg MJ, Freemont PS:
The BRCA1 C-terminal domain: structure and function.
Mutat Res 2000, 460:319-332.
34. Yu X, Chini CC, He M, Mer G, Chen J: The BRCT domain is a
phospho-protein binding domain. Science 2003, 302:639-642.
35. Xu X, Lee J, Stern DF: Microcephalin is a DNA damage response


protein involved in regulation of CHK1 and BRCA1. J Biol Chem
2004, 279:34091-34094.
Epitope-tagged microcephalin proteins were observed by the authors to
localise to ionising radiation-induced foci, suggesting a role in DNA
damage repair. Further work using RNA interference-mediated inhibition
of microcephalin expression, found that depletion of microcephalin led to
reduced levels of Brca1 and Chk1, and also defective intra-S and G2/M
checkpoints after ionising radiation.
36. Lin SY, Elledge SJ: Multiple tumor suppressor pathways

negatively regulate telomerase. Cell 2003, 113:881-889.
Microcephalin was isolated in a cellular screen for negative regulators of
hTERT transcription. The authors called the gene BRIT1 (BRCT-repeat
inhibitor of hTERT expression) and demonstrated that small interfering
RNA of BRIT1 (microcephalin) could derepress hTERT expression in
U2OS cells.
37. Gao Y, Sun Y, Frank KM, Dikkes P, Fujiwara Y, Seidl KJ,
Sekiguchi JM, Rathbun GA, Swat W, Wang J et al.: A critical role
for DNA end-joining proteins in both lymphogenesis and
neurogenesis. Cell 1998, 95:891-902.
38. Hakem R, de la Pompa JL, Sirard C, Mo R, Woo M, Hakem A,
Wakeham A, Potter J, Reitmair A, Billia F et al.: The tumor
suppressor gene Brca1 is required for embryonic cellular
proliferation in the mouse. Cell 1996, 85:1009-1023.
39. Evans PD, Anderson JR, Vallender EJ, Gilbert SL, Malcom CM,

Dorus S, Lahn BT: Adaptive evolution of ASPM, a major
Current Opinion in Genetics & Development 2005, 15:241248
248 Genetics of disease

determinant of cerebral cortical size in humans. Hum Mol
Genet 2004, 13:489-494.
See annotation [51

].
40. do Carmo Avides M, Glover DM: Abnormal spindle protein, Asp,
and the integrity of mitotic centrosomal microtubule
organizing centers. Science 1999, 283:1733-1735.
41. Riparbelli MG, Callaini G, Glover DM, Avides Mdo C: A
requirement for the abnormal spindle protein to organise
microtubules of the central spindle for cytokinesis in
Drosophila. J Cell Sci 2002, 115:913-922.
42. Gonzalez C, Saunders RD, Casal J, Molina I, Carmena M, Ripoll P,
Glover DM: Mutations at the asp locus of Drosophila lead to
multiple free centrosomes in syncytial embryos, but restrict
centrosome duplication in larval neuroblasts. J Cell Sci 1990,
96:605-616.
43. Riparbelli MG, Massarelli C, Robbins LG, Callaini G: The abnormal
spindle protein is required for germ cell mitosis and oocyte
differentiation during Drosophila oogenesis. Exp Cell Res 2004,
298:96-106.
44. Kraut R, Chia W, Jan LY, Jan YN, Knoblich JA: Role of
inscuteable in orienting asymmetric cell divisions in
Drosophila. Nature 1996, 383:50-55.
45. Chenn A, McConnell SK: Cleavage orientation and the
asymmetric inheritance of Notch1 immunoreactivity in
mammalian neurogenesis. Cell 1995, 82:631-641.
46. Wang Y-q, Su B: Molecular evolution of microcephalin, a gene

determining human brain size. Hum Mol Genet 2004,
13:1131-1137.
In addition to demonstrating rapid evolution of microcephalin in the
common ancestor of great apes, these authors predict four substitutions
that have arisen by positive selection in the human population.
47. Springer MS, Murphy WJ, Eizirik E, OBrien SJ: Placental
mammal diversification and the CretaceousTertiary
boundary. Proc Natl Acad Sci USA 2003, 100:1056-1061.
48. Russo CA, Takezaki N, Nei M: Molecular phylogeny and
divergence times of drosophilid species. Mol Biol Evol 1995,
12:391-404.
49. Manke IA, Lowery DM, Nguyen A, Yaffe MB: BRCT repeats as
phosphopeptide-binding modules involved in protein
targeting. Science 2003, 302:636-639.
50. Zhang J: Evolution of the human ASPM gene, a major

determinant of brain size. Genetics 2003, 165:2063-2070.
See annotation [51

].
51. Kouprina N, Pavlicek A, Mochida GH, Solomon G, Gersch W,


Yoon YH, Collura R, Ruvolo M, Barrett JC, Woods CG et al.:
Accelerated evolution of the ASPM gene controlling brain
size begins prior to human brain expansion. PLoS Biol 2004,
2:126.
This study, together with that of Evans et al. [39
] and Zhang [50
], identifies
the signatures of strong positive selection in the ASPM gene of great apes.
Impressively, the authors sequence and analyse complete primate ASPM
genes including their promoters, non-coding and coding sequences.
52. Pavlicek A, Noskov VN, Kouprina N, Barrett JC, Jurka J,
Larionov V: Evolution of the tumor suppressor BRCA1 locus
in primates: implications for cancer predisposition. Hum Mol
Genet 2004, 13:2737-2751.
53. Rehen SK, McConnell MJ, Kaushal D, Kingsbury MA, Yang AH,
Chun J: Chromosomal variation in neurons of the developing
and adult mammalian nervous system. Proc Natl Acad Sci USA
2001, 98:13361-13366.
54. Evolutionary history of a gene controlling brain size. PLoS Biol
2004, 2:e134.

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