A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype

Pamela Magini; Tommaso Pippucci; I. Chun Tsai; Simona Coppola; Emilia Stellacci; Anna Bartoletti-Stella; Daniela Turchetti; Claudio Graziano; Giovanna Cenacchi; Iria Neri; Duccio Maria Cordelli; Valentina Marchiani; Rosalba Bergamaschi; Giuseppe Gasparre; Giovanni Neri; Laura Mazzanti; Annalisa Patrizi; Emilio Franzoni; Giovanni Romeo; Domenico Bordo; Marco Tartaglia; Elias Nicholas Katsanis; Marco Seri

doi:10.1093/hmg/ddu070

A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype

Pamela Magini, Tommaso Pippucci, I. Chun Tsai, Simona Coppola, Emilia Stellacci, Anna Bartoletti-Stella, Daniela Turchetti, Claudio Graziano, Giovanna Cenacchi, Iria Neri, Duccio Maria Cordelli, Valentina Marchiani, Rosalba Bergamaschi, Giuseppe Gasparre, Giovanni Neri, Laura Mazzanti, Annalisa Patrizi, Emilio Franzoni, Giovanni Romeo, Domenico BordoMarco Tartaglia, Elias Nicholas Katsanis, Marco Seri^*

^*Corresponding author for this work

Pediatrics

Research output: Contribution to journal › Article › peer-review

36 Scopus citations

Abstract

Loss-of-function mutations in PAK3 contribute to non-syndromic X-linked intellectual disability (NS-XLID) by affecting dendritic spine density and morphology. Linkage analysis in a three-generation family with affected males showing ID, agenesis of corpus callosum, cerebellar hypoplasia, microcephaly and ichthyosis, revealed a candidate disease locus in Xq21.33q24 encompassing over 280 genes. Subsequent to sequencing all coding exons of the X chromosome, we identified a single novel variant within the linkage region, affecting a conserved codon of PAK3. Biochemical studies showed that, similar to previous NS-XLID-associated lesions, the predicted amino acid substitution (Lys389Asn) abolished the kinase activity of PAK3. In addition, the introduced residue conferred a dominant-negative function to the protein that drives the syndromic phenotype. Using a combination of in vitro and in vivo studies in zebrafish embryos,weshow that PAK3^N389 escapes its physiologic degradation and is able to perturb MAPK signaling via an uncontrolled kinase-independent function, which in turn leads to alterations of cerebral and craniofacial structures in vivo. Our data expand the spectrum of phenotypes associated with PAK3 mutations, characterize a novel mechanism resulting in a dual molecular effect of the same mutation with a complex PAK3 functional deregulation and provide evidence for a direct functional impact of aberrant PAK3 function on MAPK signaling.

Original language	English (US)
Article number	ddu070
Pages (from-to)	3607-3617
Number of pages	11
Journal	Human molecular genetics
Volume	23
Issue number	13
DOIs	https://doi.org/10.1093/hmg/ddu070
State	Published - Jul 2014

Funding

Medical and family history, with particular regard to neurological and cutaneous involvement, was obtained. Physical examination of the proband III-2 and of his cousin III-4 included assessment of height, weight and dysmorphic features. The neurological examination was especially focused on the evaluation of the electric activity of the brain by EEG and on the detection of cerebral malformations by MRI. The dermatological evaluation was supported by skin biopsy and was extended to carrier females. After abortion, the fetus III-5 was subjected to autopsy to confirm the brain anomaly previously detected by ultrasound scan and to reveal possible skin alterations.

ASJC Scopus subject areas

Molecular Biology
Genetics
Genetics(clinical)

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Magini, P., Pippucci, T., Tsai, I. C., Coppola, S., Stellacci, E., Bartoletti-Stella, A., Turchetti, D., Graziano, C., Cenacchi, G., Neri, I., Cordelli, D. M., Marchiani, V., Bergamaschi, R., Gasparre, G., Neri, G., Mazzanti, L., Patrizi, A., Franzoni, E., Romeo, G., ... Seri, M. (2014). A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype. Human molecular genetics, 23(13), 3607-3617. Article ddu070. https://doi.org/10.1093/hmg/ddu070

@article{3a90b3a44c2946e7a3bdaacc1855f7de,

title = "A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype",

abstract = "Loss-of-function mutations in PAK3 contribute to non-syndromic X-linked intellectual disability (NS-XLID) by affecting dendritic spine density and morphology. Linkage analysis in a three-generation family with affected males showing ID, agenesis of corpus callosum, cerebellar hypoplasia, microcephaly and ichthyosis, revealed a candidate disease locus in Xq21.33q24 encompassing over 280 genes. Subsequent to sequencing all coding exons of the X chromosome, we identified a single novel variant within the linkage region, affecting a conserved codon of PAK3. Biochemical studies showed that, similar to previous NS-XLID-associated lesions, the predicted amino acid substitution (Lys389Asn) abolished the kinase activity of PAK3. In addition, the introduced residue conferred a dominant-negative function to the protein that drives the syndromic phenotype. Using a combination of in vitro and in vivo studies in zebrafish embryos,weshow that PAK3N389 escapes its physiologic degradation and is able to perturb MAPK signaling via an uncontrolled kinase-independent function, which in turn leads to alterations of cerebral and craniofacial structures in vivo. Our data expand the spectrum of phenotypes associated with PAK3 mutations, characterize a novel mechanism resulting in a dual molecular effect of the same mutation with a complex PAK3 functional deregulation and provide evidence for a direct functional impact of aberrant PAK3 function on MAPK signaling.",

author = "Pamela Magini and Tommaso Pippucci and Tsai, {I. Chun} and Simona Coppola and Emilia Stellacci and Anna Bartoletti-Stella and Daniela Turchetti and Claudio Graziano and Giovanna Cenacchi and Iria Neri and Cordelli, {Duccio Maria} and Valentina Marchiani and Rosalba Bergamaschi and Giuseppe Gasparre and Giovanni Neri and Laura Mazzanti and Annalisa Patrizi and Emilio Franzoni and Giovanni Romeo and Domenico Bordo and Marco Tartaglia and Katsanis, {Elias Nicholas} and Marco Seri",

note = "Funding Information: Medical and family history, with particular regard to neurological and cutaneous involvement, was obtained. Physical examination of the proband III-2 and of his cousin III-4 included assessment of height, weight and dysmorphic features. The neurological examination was especially focused on the evaluation of the electric activity of the brain by EEG and on the detection of cerebral malformations by MRI. The dermatological evaluation was supported by skin biopsy and was extended to carrier females. After abortion, the fetus III-5 was subjected to autopsy to confirm the brain anomaly previously detected by ultrasound scan and to reveal possible skin alterations.",

year = "2014",

month = jul,

doi = "10.1093/hmg/ddu070",

language = "English (US)",

volume = "23",

pages = "3607--3617",

journal = "Human molecular genetics",

issn = "0964-6906",

publisher = "Oxford University Press",

number = "13",

}

Magini, P, Pippucci, T, Tsai, IC, Coppola, S, Stellacci, E, Bartoletti-Stella, A, Turchetti, D, Graziano, C, Cenacchi, G, Neri, I, Cordelli, DM, Marchiani, V, Bergamaschi, R, Gasparre, G, Neri, G, Mazzanti, L, Patrizi, A, Franzoni, E, Romeo, G, Bordo, D, Tartaglia, M, Katsanis, EN & Seri, M 2014, 'A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype', Human molecular genetics, vol. 23, no. 13, ddu070, pp. 3607-3617. https://doi.org/10.1093/hmg/ddu070

TY - JOUR

T1 - A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype

AU - Magini, Pamela

AU - Pippucci, Tommaso

AU - Tsai, I. Chun

AU - Coppola, Simona

AU - Stellacci, Emilia

AU - Bartoletti-Stella, Anna

AU - Turchetti, Daniela

AU - Graziano, Claudio

AU - Cenacchi, Giovanna

AU - Neri, Iria

AU - Cordelli, Duccio Maria

AU - Marchiani, Valentina

AU - Bergamaschi, Rosalba

AU - Gasparre, Giuseppe

AU - Neri, Giovanni

AU - Mazzanti, Laura

AU - Patrizi, Annalisa

AU - Franzoni, Emilio

AU - Romeo, Giovanni

AU - Bordo, Domenico

AU - Tartaglia, Marco

AU - Katsanis, Elias Nicholas

AU - Seri, Marco

N1 - Funding Information: Medical and family history, with particular regard to neurological and cutaneous involvement, was obtained. Physical examination of the proband III-2 and of his cousin III-4 included assessment of height, weight and dysmorphic features. The neurological examination was especially focused on the evaluation of the electric activity of the brain by EEG and on the detection of cerebral malformations by MRI. The dermatological evaluation was supported by skin biopsy and was extended to carrier females. After abortion, the fetus III-5 was subjected to autopsy to confirm the brain anomaly previously detected by ultrasound scan and to reveal possible skin alterations.

PY - 2014/7

Y1 - 2014/7

N2 - Loss-of-function mutations in PAK3 contribute to non-syndromic X-linked intellectual disability (NS-XLID) by affecting dendritic spine density and morphology. Linkage analysis in a three-generation family with affected males showing ID, agenesis of corpus callosum, cerebellar hypoplasia, microcephaly and ichthyosis, revealed a candidate disease locus in Xq21.33q24 encompassing over 280 genes. Subsequent to sequencing all coding exons of the X chromosome, we identified a single novel variant within the linkage region, affecting a conserved codon of PAK3. Biochemical studies showed that, similar to previous NS-XLID-associated lesions, the predicted amino acid substitution (Lys389Asn) abolished the kinase activity of PAK3. In addition, the introduced residue conferred a dominant-negative function to the protein that drives the syndromic phenotype. Using a combination of in vitro and in vivo studies in zebrafish embryos,weshow that PAK3N389 escapes its physiologic degradation and is able to perturb MAPK signaling via an uncontrolled kinase-independent function, which in turn leads to alterations of cerebral and craniofacial structures in vivo. Our data expand the spectrum of phenotypes associated with PAK3 mutations, characterize a novel mechanism resulting in a dual molecular effect of the same mutation with a complex PAK3 functional deregulation and provide evidence for a direct functional impact of aberrant PAK3 function on MAPK signaling.

AB - Loss-of-function mutations in PAK3 contribute to non-syndromic X-linked intellectual disability (NS-XLID) by affecting dendritic spine density and morphology. Linkage analysis in a three-generation family with affected males showing ID, agenesis of corpus callosum, cerebellar hypoplasia, microcephaly and ichthyosis, revealed a candidate disease locus in Xq21.33q24 encompassing over 280 genes. Subsequent to sequencing all coding exons of the X chromosome, we identified a single novel variant within the linkage region, affecting a conserved codon of PAK3. Biochemical studies showed that, similar to previous NS-XLID-associated lesions, the predicted amino acid substitution (Lys389Asn) abolished the kinase activity of PAK3. In addition, the introduced residue conferred a dominant-negative function to the protein that drives the syndromic phenotype. Using a combination of in vitro and in vivo studies in zebrafish embryos,weshow that PAK3N389 escapes its physiologic degradation and is able to perturb MAPK signaling via an uncontrolled kinase-independent function, which in turn leads to alterations of cerebral and craniofacial structures in vivo. Our data expand the spectrum of phenotypes associated with PAK3 mutations, characterize a novel mechanism resulting in a dual molecular effect of the same mutation with a complex PAK3 functional deregulation and provide evidence for a direct functional impact of aberrant PAK3 function on MAPK signaling.

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U2 - 10.1093/hmg/ddu070

DO - 10.1093/hmg/ddu070

M3 - Article

C2 - 24556213

AN - SCOPUS:84902349968

SN - 0964-6906

VL - 23

SP - 3607

EP - 3617

JO - Human molecular genetics

JF - Human molecular genetics

IS - 13

M1 - ddu070

ER -

A mutation in PAK3 with a dual molecular effect deregulates the RAS/MAPK pathway and drives an X-linked syndromic phenotype

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