Progranulin mutations in Dutch familial frontotemporal lobar degeneration

ArticleinEuropean Journal of HumanGenetics 15(3):369-74 · April 2007
Impact Factor: 4.35 · DOI: 10.1038/sj.ejhg.5201772 · Source: PubMed
Abstract

Mutations in the progranulin (PGRN) gene have recently been identified in frontotemporal lobar degeneration with ubiquitin inclusions linked to chromosome 17q21. We report here the finding of two novel frameshift mutations and three possible pathogenic missense mutations in the PGRN gene. Furthermore, we determined the frequency of PGRN mutations in familial cases recruited from a large population-based study of frontotemporal lobar degeneration carried out in The Netherlands.

Full-text

Available from: Patrizia Rizzu, Mar 05, 2015
ARTICLE
Progranulin mutations in Dutch familial
frontotemporal lobar degeneration
Iraad F Bronner
1,2,4
, Patrizia Rizzu*
,1,2,4
, Harro Seelaar
3
, Saskia E van Mil
1,2
, Burcu Anar
1,2
,
Asma Azmani
3
, Laura Donker Kaat
3
, Sonia Rosso
3
, Peter Heutink
1,2
and John C van Swieten
3
1
Department of Human Genetics, Section Medical Genomics, VU University Medical Center and VU University,
Amsterdam, The Netherlands;
2
Center for Neurogenomics and Cognitive Research, VU University Medical Center and
VU University, Amsterdam, The Netherlands;
3
Department of Neurology, Erasmus MC University Medical Center,
Rotterdam, The Netherlands
Mutations in the progranulin (PGRN) gene have recently been identified in frontotemporal lobar
degeneration with ubiquitin inclusions linked to chromosome 17q21. We report here the finding of two
novel frameshift mutations and three possible pathogenic missense mutations in the PGRN gene.
Furthermore, we determined the frequency of PGRN mutations in familial cases recruited from a large
population-based study of frontotemporal lobar degeneration carried out in The Netherlands.
European Journal of Human Genetics (2007) 15, 369374. doi:10.1038/sj.ejhg.5201772; published online 17 January 2007
Keywords: PGRN; FTLD; mutations; neurodegeneration
Introduction
The term frontotemporal lobar degeneration (FTLD) refers
to an heterogeneous group of neurodegenerative disorders
clinically characterized by progressive behavioral changes
and cognitive dysfunctions, including executive and
language functions.
1
Sometimes, language impairment
presents as an initial symptom sub-classifying this FTLD
group into progressive nonfluent aphasia and semantic
dementia. Additionally, the clinical picture can be compli-
cated by motor symptoms such as motor neuron disease
(MND) or parkinsonism.
2
Two main pathological FTLD subtypes are recognized
based on the presence of tau-positive inclusions (tauopa-
thies) or tau-negative ubiquitin-positive neuronal inclu-
sions (FTLD-U).
3
Characteristically the ubiquitin
immunoreactive inclusions (ub-i) are observed in the
dentate gyrus of the hippocampus and in the superficial
layers of the frontal and temporal cortex.
4
A positive family history is found in approximately 40%
of FTLD cases, and linkage studies have shown that FTLD is
genetically heterogeneous with loci and genes identified
on chromosomes 3 (FTD3),
5
9p,
6
9q
7
and 17q (FTDP-17
8
and FTDU-17
9,10
). Recently, mutations in the PGRN gene
were found in several families with FTDU-17.
9,10
PGRN
encodes a biologically active precursor glycoprotein de-
scribed previously as a multifunctional growth factor
involved in development, inflammation and wound
repair.
11
In the present study, we report the finding of two novel
frameshift mutations and three possible pathogenic mis-
sense mutations in the progranulinv (PGRN) gene. In
addition, we describe the genetic contribution of PGRN
to FTLD in a series of familial cases recruited from a large
cohort of FTLD patients.
Materials and methods
Patients
Three hundred and thirty-eight patients with FTLD (182
females and 156 males) with mean age at onset of
Received 25 September 2006; revised 29 November 2006; accepted 30
November 2006; published online 17 January 2007
*Correspondence: Dr P Rizzu, Department of Human Genetics, Section
Medical Genomics and Center for Neurogenomics and Cognitive
Research, VU University Medical Center and VU University, Amsterdam,
van der Boechorststraat 7, 1081 BT Amsterdam, The Netherlands.
Tel: þ 31 20 5989961; Fax: þ 31 20 4448285; E-mail: p.rizzu@vumc.nl
4
These authors contributed equally to this work.
European Journal of Human Genetics (2007) 15, 369374
&
2007 Nature Publishing Group All rights reserved 1018-4813/07
$30.00
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Page 1
57.479.3 years were identified in a genetic epidemio-
logical study in the Netherlands. The clinical diagnosis in
all patients was established according to international
consensus criteria.
12
Clinical family history was positive in
166 patients (59%) and among them DNA was available in
137 cases. Eighty-seven of these 137 patients came from
independent families: 10 families presented MAPT muta-
tions,
13
two large families showed FTLD-U with definite
linkage to chromosome 17q2122, six smaller families had
multiple (42) affecteds and 69 had two affecteds.
DNA study
The 13 exons of PGRN including intron/exons boundaries
were amplified from genomic DNA by PCR and directly
sequenced in both strands. Novel sequence variants were
analyzed in a minimum of 380 chromosomes from healthy
individuals of matched ethnicity.
Immunohistochemistry
Immunohistochemistry experiments were performed on
eight available brains as described previously.
14
Results
To determine the possible involvement of the newly found
PGRN gene in our cohort, we systematically screened for
mutations in 77 cases with positive family history of
dementia consistent with autosomal dominant pattern of
inheritance and with no MAPT and CHMP2B mutations.
The mean age at onset in this group was 59.379.1 years.
We identified two novel frameshift mutations Ser82-
Valfs174X and Val411Sfr1X (Table 1) predicted to cause
premature termination of the coding sequence likely
leading to loss of functional PGRN protein similar to
previous reports. One nonsense mutation (Gln125X) was
also observed in a independently ascertained member of
the 1083 FTLD-U family already described.
10
Furthermore,
we identified five novel coding sequence variants (three
missense and two silent mutations), two intronic sequence
changes in intron 2 and 7 and the previously reported
missense mutation Gly414Val.
15
The frameshift mutations
and the GGG93GGA, Thr182Met, Pro233His, CAC447CAT
and Trp541Cys mutations were not found in controls;
in contrast, the two intronic variants were also present in
healthy individuals suggesting they are not pathogenic.
Moreover, the GGG93GGA silent mutation was detected in
co-occurrence with the Pro233His.
The Ser82Valfs174X mutation was found in a 69-year-old
woman, member of the HFTD3 family previously linked to
17q2122
4
(Figure 1a). A large variation in age at onset
(between 45 and 75 years) was observed between affected
individual from this family.
Sequencing of 13 additional DNA samples from affected
family members showed complete segregation of the
mutation with the disease. The clinical symptoms in this
family consisted of apathy, loss of initiative and interest,
roaming behaviour and word finding difficulties. Two
patients developed parkinsonism early in the course of
the disease, which moderately responded to levodopa
treatment. Signs of motor neuron disease were not
observed. Extensive neuropsychological testing in six
patients revealed impaired naming with normal compre-
hension of language.
The Val411Serfr1X mutation was identified in a 66-year-
old woman, who presented with speech, and writing errors,
and word finding difficulties. The patient showed social
inappropriate behaviour and emotional bluntness. Magnetic
resonance imaging showed asymmetric right-sided fronto-
temporal atrophy. The patient developed loss of initiative,
and died from bronchopneumonia. Her mother and grand-
mother suffered from identical symptoms, whereas her uncle
and a nephew were diagnosed as Pick’s disease.
Table 1 PGRN mutations identified in FTLD patients and healthy control individuals
Location Genomic
a
Predicted cDNA
b
Protein
c
Rs number Patients (N) Controls (N)
Exon 2 g.4407delC c.243delC Ser82ValfsX174 HFTD3 F
Intron 2 g.4436G4A 14
Intron 2 g.4445G4A rs9897526 19 35
Exon 3 g.4559G4A c.279G4A Gly93Gly 1 F
Intron3 g.4661G4C F 1
Exon 4 g.5129C4T c.592C4T Gln125X 1 F
Exon 5 g.5402C4T c.545C4T Thr182Met 1 F
Exon 6 g.56674A c.698C4A Pro233His 1 F
Intron 7 g.6048G4A 22 18
Exon 10 g.6944_6945 delGT c.1231_1232delGT Val411SerfsX1 1 F
Exon10 g.6954G4T c.1241G4T Gly414Val 1 1
Exon10 g.7054C4T c.1341C4T His447His 1 F
Exon10 g.6966G4A c.1253G4A Arg418Gln F 2
Exon11 g.7428G4C c.1623G4C Trp541Cys 1 F
a
Numbering relative to NC_000017.9 Genbank Accession Number and starting at nucleotide 1.
b
Numbering relative to NM_002087.2 starting at the ATG.
c
Numbering relative to NP_002087.1.
PGRN mutations in FTLD
IF Bronner et al
370
European Journal of Human Genetics
Page 2
The Gln125X mutation was found in a 60-year-old
woman, who came from the family 1083, described
previously.
10
She presented with memory problems and
word findings difficulties.
Neuropathological examination showed ub-i in dentate
gyrus, neocortex and/or striatum (Figure 1b and c).
Ubiquitin inclusions were also present in six additional
brains from FTLD cases with no PGRN mutations with the
distinct morphology and distribution pattern characteristic
of FTLD-U type 2,
16
including several neuronal intra-
nuclear inclusions in the frontal neocortex in one case
(Figure 1d).
Discussion
The present study report the identification of three
pathogenic (Val411Serfr1X and Ser82Valfs174X as novel)
PGRN mutations that account for B4% of the independent
familial FTLD cases.
Similar to previous PGRN studies the two novel muta-
tions determine a frameshift, which results in the genera-
tion of premature termination codons. Eukaryotic cells
are capable to detect and degrade transcripts harbouring
premature signals for the termination of translation
through the nonsense-mediated mRNA decay (NMD)
pathway. Degradation of mutant mRNAs results in null
alleles
9,15
with loss of functional PGRN.
Several rare missense and silent mutations were detected
in patients but not in controls. Segregation studies could
not be performed in these cases, as DNA from affected
family members was not available.
Although it cannot be excluded that these changes are
benign variants as they are located in granulin domains
each composed of 7,5 tandem repeats of highly conserved
motifs of 12 cystein residues suggested to be functional
redundant,
15
several studies have shown that separate
repeats may have alternative binding capacities and there-
fore different functions,
17
highlighting the possibility that
these variants are pathogenic. The Pro233 and the Trp541,
in particular, are highly conserved among species
(Figure 2a) and in the granulin domains (Figure 2b).
Furthermore, previous reports have suggested these
amino acids are essential for the proper folding of the
protein.
18 20
The Trp residue is likely involved in the
hydrophobic packaging of the beta-sheet and substitution
with the cys residue, which has the ability to form
disulfide bridges, might affect PGRN 3D structure. The
Pro residue is part of an antiparallel beta-sheet, and
might be important for stacking multiple repeats,
Figure 1 Pedigree and ubiquitin pathology (a). Pedigree of family HFTD3; only affected individuals are shown. This large family consists of 42
affected and 102 unaffected members. Arrow indicates a healthy carrier: a 72-year-old mother of two affected sisters who carried the mutation and did
not have any cognitive complaints and behavioural changes as confirmed by reports of other family members and by neurological examination. (b d)
Ubiquitin staining; ubiquitin-positive neuronal cytoplasmatic inclusions in granular cells of the dentate gyrus (b) and in the superficial layers of the
frontal neocortex (c) with ubiquitin-positive dystrophic neurites (c) in Ser82Valfs174X brain. Lentiform ubiquitin-positive neuronal intranuclear
inclusion in one FTLD case with no PGRN mutations (d). Scale bars: 100 mm.
PGRN mutations in FTLD
IF Bronner et al
371
European Journal of Human Genetics
Page 3
GRANULIN MOTIF II
1 HOMO SAPIENS IQCPDSQFECPDFST CCVMVDGSWGCCPMP QASCCEDRVHCCPHG AFCDLVHTRCITPT- -GTHPLAKKLPAQRT NRAV----------- 200
2 PAN TROGLODYTES --------------- --------------- -ASCCEDRVHCCPHG AFCDLVHTRCITPT- -GTHPLAKKLPAQRT NRAV----------- 46
3 MACACA MULATTA VQCPDSHFECPDLST CCVMVDGSWGCCPMP QASCCEDRVHCCPHG AFCDLVHTRCITPT- -GTHPLAKKIPAQRS NRAV----------- 200
4 BOS TAURUS VQCPDKQFQCPNSST CCTMPDGSWGCCPMP QASCCEDKIHCCPHG TSCDLARGRCLSAT- -GTHPLAKKMPAHKT KSSA----------- 213
5 CANIS FAMILIARIS IQCPDSQFLCPNSST CCTMLDGSWGCCPMP QASCCEDKVHCCPHG TSCDLAHARCLTAT- -GSHPLAKKIPAQRS NR------------- 199
6 RATTUS NORVEGICUS VQCPGSQFECPDSAT CCIMIDGSWGCCPMP QASCCEDRVHCCPHG ASCDLVHTRCISPT- -GTHPLLKKFPAQRT NRAV----------- 199
7 MUS MUSCULUS VQCPGSQFECPDSAT CCIMVDGSWGCCPMP QASCCEDRVHCCPHG ASCDLVHTRCVSPT- -GTHTLLKKFPAQKT NRAV----------- 212
8 DASYPUS NOVEMCINCTU IQCPDSQFECPDFST CCVMVDGSWGCCPMP QASCCEDRVHCCPHG AFCDLVHTRCITPT- -GTHPLAKKLPAQRT NRAV----------- 200
9 MONODELPHIS DOMESTICA VKCPDSEFECPDEST CCMMQDGSWGCCPMP KASCCEDKVHCCPQG SVCDLAHSRCLTSG- -GTYPLAQKNPAKKI QQEKD---------- 201
10 DANIO RERIO GRNA DVACNDTAACPDGST CCKTKDGGWACCPLP EAVCCEDFIHCCPHG KKCDVAAGSCEDPS- -GSVSWVEKVPVRPI RKQK----------- 364
11 DANIO RERIO GRNB VICPDKISKCPEDTT CCLLETGSYGCCPMP KAVCCSDQKHCCPEG TTCDLIHSTCLSAN- -GVSEMAIKIPAVT- --------------- 256
12 TAKIFUGU RUBRIPES IICPDGKSRCQLGHT CCQLASGAYGCCPLQ QAVCCSDHERCCPAG TRCDLEHDACVSG-- -ATPVPMLRIAAVPG EGTRVFAAT--AAET 256
13 TETRAODON NIGROVIRIDIS VICPDGKSSCSEGAT CCQLTSGEYGCCPYP QAVCCSDHLHCCPTG TRCDLALSVCVAGP- -GGPSPASKIIAALG PEPKSSGQV--FAGV 215
14 GASTEROSTEUS ACULEATUS GRNA VLCKDGVSECPDGTT CCENPDGKWACCPLP KAVCCEDKTHCCPEG TTCDVEHSKCISLFT KQELPMWAKSPARLR ADWENPKDRFPLSEQ 259
15 GASTEROSTEUS ACULEATUS GRNB VSCPGGKSSCPDSYT CCLLASGDYGCCPYS QAMCCSDHLHCCPSN TICDLAHGVCKDGE- -AIFPLLKKIAAVPN DVTCPDETS--SCPD 264
16 CIONA INTESTINALIS VQCPDGRSACPDGNT CCKLASGAYGCCPQP KAVCCSDHVHCCPQG YSCNVGSGTCLKQDS LSVVPWMEKQEAVTL NVGMVQCPDGHSACP 331
GRANULIN MOTIF III
1 HOMO SAPIENS -------------AL S-SSVMCPDARSRCP DGSTCCELPSGKYGC CPMPNATCCSDHLHC CPQDTVCDLIQSKCL SKENATTDLLTKLPA 276
2 PAN TROGLODYTES -------------AL S-SSVMCPDARSQCP DGSTCCELPSGKYGC CPMPNATCCSDHLHC CPQDTVCDLVQSKCL SKENATTDLLTKLPA 122
3 MACACA MULATTA -------------AL S-SSVMCPDARSQCP DGSTCCELPSGKYGC CPMPNAMCCSDHLHC CPQDTVCDLIQSKCL SKENTTMDLLTKLPA 276
4 BOS TAURUS -------------FF PLPVILCPDGQSQCP DGSTCCKLPTGKYGC CPMPNAICCSDHLHC CPQNTVCDLTQSKCL SKE-NATDLLTKLPA 289
5 CANIS FAMILIARIS --------------- --AGVICPDGRSQCP DGSTCCELPSGKYGC CPMPHAICCSDHLHC CPQDTVCDLVRSKCL SKE-NATDLLTKLPA 271
6 RATTUS NORVEGICUS -------------AF P-FSVVCPDAKTQCP DDSTCCELPTGKYGC CPMPNAICCSDHLHC CPQDTVCDLIQSKCI SKD-YTTDLMTKLPG 274
7 MUS MUSCULUS -------------SL P-FSVVCPDAKTQCP DDSTCCELPTGKYGC CPMPNAICCSDHLHC CPQDTVCDLIQSKCL SKN-YTTDLLTKLPG 287
8 DASYPUS NOVEMCINCTUS -------------AL S-SSVMCPDARSRCP DGSTCCELPSGKYGC CPMPNATCCSDHLHC CPQDTVCDLIQSKCL SKENATTDLLTKLPA 276
9 MONODELPHIS DOMESTICA -------------VT VTTNRLCPDGRSQCS DGTTCCQLPSGSYGC CPLPNAICCPDHMHC CPQNTVCDLEKSECL SKNGSASGLFVKLPA 278
10 DANIO RERIO GRNA --------VAVTQVS SVSSDVPCNDTAACA DGTTCCKTKEGDWAC CPLPEAVCCEDFVHC CPKGKKCNIAAMKCE DPLCTEEPLVKQTPV 446
11 DANIO RERIO GRNB ------------VLK PKEEVVPCNETVACS SGTTCCKTPEGSWAC CPLPKAVCCEDHIHC CPEGTLCNVAASSCD DPTELSVSVPWMEKV 334
12 TAKIFUGU RUBRIPES ---------ATPVVP IKIDNNKCDESTTCP GDSTCCRTLEGGWAC CPLAQAVCCDDHVHC CPHDTICNLETQTCD GQSGGRPPLRWVEKV 353
13 TETRAODON NIGROVIRIDIS ----SPGVATTPVLP FLPDDTKCDDTASCP GDYTCCRTLKGGWAC CPLAQAVCCSDHTHC CPHNTICNLQERTCN SQSGGRPPLRWVEKV 318
14 GASTEROSTEUS ACULEATUS GRNA VPRPGFRHEGNFAVV FAGVSVACDATEACA GNSTCCMTPEGGWSC CPLPEAVDCEDSVHC CPKGRKCNPATQACD SEGCSVPWLQKVPTI 439
15 GASTEROSTEUS ACULEATUS GRNB AALTVVAVATPTEVQ KKVSVLPCNDSVACA EGSTCCGLVEGGWAC CPLPKAVCCEDHQHC CPHGTVCDLEASTCV DSSAG-TSTPWFDKS 402
16 CIONA INTESTINALIS LTVVPWMEKQDSVAF NVGMVQCPDGRSACP DGNTCCKLASGAYGC CPQPKAVCCSDHVHC CPQGYSCNVGSGTCL KQD-SLSVVPWMEKQ 469
GRANULIN MOTIF V-a
1 HOMO SAPIENS LSLPD---------- --PQ----ALKRDVP CDNVSSCPSSDTCCQ LTSG-EWGCCPIP-- --------------- --------------- 392
2 PAN TROGLODYTES LSLPD---------- --PQ----ALKRDVP CDNVSSCPSSDTCCQ LMSG-EWGCCPIP-- --------------- --------------- 238
3 MACACA MULATTA LSLPD---------- --PQ----ALKRDVP CHIVS---------- --------------- --------------- --------------- 370
4 BOS TAURUS LSLLD---------- --LG----AVEGDVP CDNVTSCPSSTTCCR LKSG-EWACCPAP-- --------------- --------------- 405
5 CANIS FAMILIARIS LQLLN---------- --PQ----ATENDVP CDNVTSCPSSNTCCR LMSG-EWGCCPAP-- --------------- --------------- 387
6 RATTUS NORVEGICUS LSLPD---------- --PQ----ILKNDVP CDDFSSCPSNNTCCR LSSG-DWGCCPIP-- --------------- --------------- 390
7 MUS MUSCULUS LRLPD---------- --PQ----ILKSDTP CDDFTRCPTNNTCCK LNSG-DWGCCPIP-- --------------- --------------- 403
8 DASYPUS NOVEMCINCTUS LSLPD---------- --PQ----ALKRDVP CDNVSSCPSSDTCCQ LTSG-EWGCCPIP-- --------------- --------------- 392
9 MONODELPHIS DOMESTICA LAAVG---------- ------------NVP CDNTTSCPSETTCCV LESG-AWGCCPAP-- --------------- --------------- 388
10 DANIO RERIO GRNA PIRKQKV-------- --AVTTASSASSDVP CNDTAACPDGSTCCK TKDG-GWACCPLP-- --------------- --------------- 578
11 DANIO RERIO GRNB ATR------------ ---AVAMPTLPARNM CDAQTSCPRDTTCCF MDQTRKWGCCPLP-- --------------- --------------- 525
12 TAKIFUGU RUBRIPES QEEPS---------- --GAASGPARPAGVM CDDRTSCPRDTSCCF MQETRRWGCCPVP-- --------------- --------------- 476
13 TETRAODON NIGROVIRIDIS VEP------------ ---SPALPAQLAEVV CDNQTSCPSHTTCCF VQKWQKFGCCPVPNV GPAVQRHRRRGQALT F-------------- 459
14 GASTEROSTEUS ACULEATUS GRNA VTAGPFPQSRATITK GEEATKAPEEDEVVQ CDSRTSCPQSNTCCF MAESQKWGCCPLPKT LCHTHCSHCRIDRFC TNHISAVKQPAEISD 606
15 GASTEROSTEUS ACULEATUS GRNB ATR------------ ---AVAMPTLPARNM CDAQTSCPRDTTCCF MDQTRKWGCCPLP-- --------------- --------------- 525
16 CIONA INTESTINALIS TFN------------ ---------VGMVQC PDGRSACPDGNTCCK LASG-AYGCCPQP-- --------------- --------------- 587
GRANULIN MOTIF V-b
1 HOMO SAPIENS --------------- --------------- --------------- ---EAVCCSDHQHCC PQGYTCVAEGQ-CQR –GSEIVA-GLEKMPA 431
2 PAN TROGLODYTES --------------- --------------- --------------- ---EAVCCSDHQHCC PQGYTCVAEGQ-CQR -GSEIVA-GLEKMPA 277
3 MACACA MULATTA --------------- --------------- --------------- --------------- --------------- --------------- 370
4 BOS TAURUS --------------- --------------- --------------- ---EAVCCSDHQHCC PKGYTCVARRH-CKR -GKQVVT-GLDKVPA 444
5 CANIS FAMILIARIS --------------- --------------- --------------- ---EAVCCSDHQHCC PHGYTCLDDGH-CQR -GSKVVS-GLEKMPA 426
6 RATTUS NORVEGICUS --------------- --------------- --------------- ---EAVCCLDHQHCC PQGFKCMDEGY-CQK -GDRMVA-GLEKMPV 429
7 MUS MUSCULUS --------------- --------------- --------------- ---EAVCCSDNQHCC PQGFTCLAQGY-CQK -GDTMVA-GLEKIPA 442
8 DASYPUS NOVEMCINCTUS --------------- --------------- --------------- ---EAVCCSDHQHCC PQGYTCVAEGQ-CQR -GSEIVA-GLEKMPA 431
9 MONODELPHIS DOMESTICA --------------- --------------- --------------- ---QAVCCPDHKHCC PHGFVCSPDG--CKS -GQKAVP-WLEKIAA 426
10 DANIO RERIO GRNA --------------- --------------- --------------- ---EAVCCEDFIHCC PHGKKCNVAAGSCDD --PSGSVPWVEKVPV 618
11 DANIO RERIO GRNB --------------- --------------- --------------- ---QAVCCADQEHCC PQGYTCDLAQSSCVR -SGLPSMAWFRKEPA 495
12 TAKIFUGU RUBRIPES --------------- --------------- --------------- ---NAVCCEDGDHCC PRGHRCDPHRRSCSK -GPLVTP-WFTKLSA 516
13 TETRAODON NIGROVIRIDIS --------------- --------------- --------------- ARPQAVCCEGGSHCC PMGHRCDPYRSSCSK -GPLVTP-WFTKLSA 502
14 GASTEROSTEUS ACULEATUS GRNA FGNRLPFTSRTCSNT KINALRCLHSPSFEE LRNGIADVKLALPPP PPPQAVCCSDGNHCC PTDYTCDVEKTTCTK G--EVVIPWYTKLPA 694
15 GASTEROSTEUS ACULEATUS GRNB --------------- --------------- --------------- ---KAVCCHDGDHCC PSGHTCEPHRSSCSK -GPLVLVPWFSKLSA 566
16 CIONA INTESTINALIS --------------- --------------- --------------- ---KAVCCSDHVHCC PQGYSCNVGSGTCLK QDSLSVVPWMEKQGE 629
GRANULIN MOTIF VII
1 HOMO SAPIENS ---------VKDVEC G--EGHFCHDNQTCC RDNRQGWACCPYRQG VCCADRRHCCPAGFR CAARGTKCLRREAPR WDAPLRDPALRQLL- 593
2 PAN TROGLODYTES ---------VKDVEC G--EGHFCHDNQTCC RDNRQGWACCPYRQG VCCADRRHCCPAGFR CAARGTKCLRREAPR WDAPLRDPALRQLL- 439
3 MACACA MULATTA -------------SW -----HFCHDNQTCC RDNRGGWACCPYRQG ICCADRRHCCPAGFR CAARGAKCLRREALR WDAPLRDPALRQLL- 441
4 BOS TAURUS ---------MGNVEC G--ARHFCHDNQTCC PDSQGGWACCPYRKG TCCGDKHHCCP---- --------------- --------------- 573
5 CANIS FAMILIARIS ---------VGNVKC G--EGHFCHDNQTCC RDSRGGWACCPYHQG VCCADQRHCCPTGFH CGAKGTKCLRRESLR WDMPLRDPAPRPLL- 588
6 RATTUS NORVEGICUS ----------GNVEC G--AGHFCHDNQSCC KDSQGGWACCPYVKG VCCRDGRHCCPIGFH CSAKGTKCLRKKTPR WDILLRDPAPRPLL- 589
7 MUS MUSCULUS ----------GNVEC G--EGHFCHDNQTCC KDSAGVWACCPYLKG VCCRDGRHCCPGGFH CSARGTKCLRKKIPR WDMFLRDPVPRPLL- 602
8 DASYPUS NOVEMCINCTUS ---------VKDVEC G--EGHFCHDNQTCC RDNRQGWACCPYRQG VCCADRRHCCPAGFR CAARGTKCLRREAPR WDAPLRDPALRQLL- 593
9 MONODELPHIS DOMESTICA ---------KGDIQC D--GKHFCHSHQTCC LARGGRWACCPLDKG VCCADGQHCCPNGFH CRAKGTKCLRRKNLR WDVLWELL------- 580
10 DANIO RERIO GRNA P-KL----DLGVVKC D--EQSSCSADSTCC LLSKDETGCCPFPEA VCCPDQKHCCPEGYR CDLRRRSCVKTTRLY VEITQLTHIRSNKPQ 786
11 DANIO RERIO GRNB -SDE----LLGHEDV KCDSSTSCPSGSTCC ILPTGQWGCCPLVKA VCCKDMKHCCPMGYK CDPKVQGCTKSSS-- --------------- 648
12 TAKIFUGU RUBRIPES RSRG----AQGVLSC GGTGEFHCPKEDTCC PTSATEWACCPSPRA VCCSDQKHCCPAGFS CDPSGG-CVQDLSSW DAWFDRS-------- 676
13 TETRAODON NIGROVIRIDIS GAGR----PEGLAPC GGTGEFHCPKEDTCC PTSATEWACCPSPGA VCCSDHKHCCPAGFS CDLKAGGCVRDPSPW DAWFHRPVRSRL--- 668
14 GASTEROSTEUS ACULEATUS GRNA AEPTSSPSEQGDVVC D--DQTSCPDGQTCC RTSATTWGCCPAPNA VCCSDMQHCCPEGHT CTETGG-CTGNNVPH WHKWQVFFSNKKRSL 865
15 GASTEROSTEUS ACULEATUS GRNB PVDH----EDVACDV G--GEFRCPGRATCC RVSASEWGCCPSPQA VCCPDSKHCCPAGYS CDPKAGGCSRPQLTW DGKSDFVPHGL---- 732
16 CIONA INTESTINALIS N--------VGMVQC P-DGRSACPDGNTCC KLASGAYGCCPQPKA VCCSDHVHCCPQGYS CNVGSGTCLKQDSLS VVPWMEKQILTKEKC 799
HUMAN PROGRANULIN
1 MWTLVSWVALTAGLVAGTRCPDGQFCPVACCLDPGGASYSCCRPLLDKWPTTLSRHLG 58
59 -GPCQVDAHCSAGHSCIFTVSGTSSCCPFPEAVACGDGHHCCPRGFHCSADGRSCFQRSGNNSVGA ---------------- 123
124 IQCPDSQFECPDFSTCCVMVDGSWGCCPMPQASCCEDRVHCCPHGAFCDLVHTRCITPTGTHPLAKKLPAQRTNRAVALSSS- 205
206 VMCPDARSRCPDGSTCCELPSGKYGCCPMPNATCCSDHLHCCPQDTVCDLIQSKCLSKENATTDLLTKLPAHTVGD------- 281
282 -VKCDMEVSCPDGYTCCRLQSGAWGCCPFTQAVCCEDHIHCCPAGFTCDTQKGTCEQGPHQVPWMEKAPAHLSLPDPQALKRD 363
464 -VPCDNVSSCPSSDTCCQLTSGEWGCCPIPEAVCCSDHQHCCPQGYTCVAEGQCQRGSEIVAGLEKMPARRASLSHPRD---- 441
542 -IGCDQHTSCPVGQTCCPSLGGSWACCQLPHAVCCEDRQHCCPAGYTCNVKARSCEKEVVSAQPATFLARSPHVGVKD----- 518
619 -VECGEGHFCHDNQTCCRDNRQGWACCPYRQGVCCADRRHCCPAGFRCAARGTKCLRREAPRWDAPLRDPALRQLL 593
CONSENSUS --C-D----CPD--TCC----G-wGCCP-----CC-D--HCCP----CD--G--C---T------------------------
a
b
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European Journal of Human Genetics
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necessary for the proper protein conformation. Therefore,
his substitution with the His residue may also change
PGRN 3D structure with consequences at functional level.
The effect of the Thr182Met mutation is less clear since it
is just outside the granulin motif. This amino acid is
conserved between mammals and was not detected in
controls.
Consistent with other PGRN studies, the clinical pre-
sentation in patients carrying the pathogenic mutations is
characterized by a large variation in age at onset and by
occurrence of symptoms of nonfluent aphasia, whereas
semantic deficits were more often seen in patients with
missense and intronic MAPT mutations within the same
cohort.
21
The HTFD3 family, in particular, shows a large
variation in age at onset. The high variability is further
confirmed by the presence of a 72-year-old healthy carrier.
Our and other findings show that a significant proportion
of patients remain unaffected until old age suggesting
therefore an interplay of several genetic and/or environ-
mental factors in the disease development.
The percentage of PGRN mutations detected in our
familial FTLD cohort (up to B7% by including the two
highly conserved missense mutations) is lower compared
to the much higher frequency observed in other studies
where PGRN mutations explain up to B25% of familial
FTLD.
10,15
The lower frequency of PGRN mutations in
our group might reflect differences in patients recruit-
ment methods, as the MAPT mutations in the Belgian
cohort account for only to 7% of all familial cases
compared to 14% detected in this cohort.
9,10,14
In
addition, in the studies by Cruts et al.
10
and Baker et al.
9
a strong founder effect among probands carrying
the IVS0 þ 5G4C and Arg493X was observed, whereas
we restricted our estimation of mutation frequency to
independent patients only.
In addition, geographical differences in frequencies may
also play a role, as seen in MAPT studies, and they cannot
be ruled out until more reports will allow a better estimate
of PGRN mutation frequency in familial FTLD.
In summary, mutations in PGRN explain only part
of FTLD in our cohort and they are absent in B80% of
cases including familial FTLD þ MND as well as FTLD-U
without MND strongly suggesting that we are only
beginning to unravel the molecular pathways leading to
FTLD and that additional genes contribute to the disease
pathogenesis.
Acknowledgements
This study was supported by the Centre for Medical Systems Biology
(CMSB), a centre of excellence approved by the Netherlands Genomics
Initiative/Netherlands Organization for Scientific Research (NWO)
and by the ‘Hersenstichting Nederland’ Project-Number 13F05(2).14
and ‘Stichting Dioraphte’.
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