Nephrocystin-1 forms a complex with polycystin-1 via a polyproline Motif/SH3 domain interaction and regulates the apoptotic response in mammals

Claas Wodarczyk, Gianfranco Distefano, Isaline Rowe, Massimiliano Gaetani, Barbara Bricoli, Mordi Muorah, Andrea Spitaleri, Valeria Mannella, Piero Ricchiuto, Monika Pema, Maddalena Castelli, Ariel E. Casanova, Luca Mollica, Manuela Banzi, Manila Boca, Corinne Antignac, Sophie Saunier, Giovanna Musco, Alessandra Boletta

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Mutations in PKD1, the gene encoding for the receptor Polycystin-1 (PC-1), cause autosomal dominant polycystic kidney disease (ADPKD). The cytoplasmic C-terminus of PC-1 contains a coiled-coil domain that mediates an interaction with the PKD2 gene product, Polycystin-2 (PC-2). Here we identify a novel domain in the PC-1 C-terminal tail, a polyproline motif mediating an interaction with Src homology domain 3 (SH3). A screen for interactions using the PC-1 C-terminal tail identified the SH3 domain of nephrocystin-1 (NPHP1) as a potential binding partner of PC-1. NPHP1 is the product of a gene that is mutated in a different form of renal cystic disease, nephronophthisis (NPHP). We show that in vitro pull-down assays and NMR structural studies confirmed the interaction between the PC-1 polyproline motif and the NPHP1 SH3 domain. Furthermore, the two full-length proteins interact through these domains; using a recently generated model system allowing us to track endogenous PC-1, we confirm the interaction between the endogenous proteins. Finally, we show that NPHP1 trafficking to cilia does not require PC-1 and that PC-1 may require NPHP1 to regulate resistance to apoptosis, but not to regulate cell cycle progression. In line with this, we find high levels of apoptosis in renal specimens of NPHP patients. Our data uncover a link between two different ciliopathies, ADPKD and NPHP, supporting the notion that common pathogenetic defects, possibly involving de-regulated apoptosis, underlie renal cyst formation.

Original languageEnglish
Article numbere12719
JournalPLoS One
Volume5
Issue number9
DOIs
Publication statusPublished - 2010

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Mammals
src Homology Domains
sequence homology
mammals
Autosomal Dominant Polycystic Kidney
apoptosis
Apoptosis
kidneys
Tail
Genes
tail
Cystic Kidney Diseases
Kidney
polyproline
polycystic kidney disease 1 protein
polycystins
Gene encoding
Cilia
genes
cilia

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Nephrocystin-1 forms a complex with polycystin-1 via a polyproline Motif/SH3 domain interaction and regulates the apoptotic response in mammals. / Wodarczyk, Claas; Distefano, Gianfranco; Rowe, Isaline; Gaetani, Massimiliano; Bricoli, Barbara; Muorah, Mordi; Spitaleri, Andrea; Mannella, Valeria; Ricchiuto, Piero; Pema, Monika; Castelli, Maddalena; Casanova, Ariel E.; Mollica, Luca; Banzi, Manuela; Boca, Manila; Antignac, Corinne; Saunier, Sophie; Musco, Giovanna; Boletta, Alessandra.

In: PLoS One, Vol. 5, No. 9, e12719, 2010.

Research output: Contribution to journalArticle

Wodarczyk, C, Distefano, G, Rowe, I, Gaetani, M, Bricoli, B, Muorah, M, Spitaleri, A, Mannella, V, Ricchiuto, P, Pema, M, Castelli, M, Casanova, AE, Mollica, L, Banzi, M, Boca, M, Antignac, C, Saunier, S, Musco, G & Boletta, A 2010, 'Nephrocystin-1 forms a complex with polycystin-1 via a polyproline Motif/SH3 domain interaction and regulates the apoptotic response in mammals', PLoS One, vol. 5, no. 9, e12719. https://doi.org/10.1371/journal.pone.0012719
Wodarczyk, Claas ; Distefano, Gianfranco ; Rowe, Isaline ; Gaetani, Massimiliano ; Bricoli, Barbara ; Muorah, Mordi ; Spitaleri, Andrea ; Mannella, Valeria ; Ricchiuto, Piero ; Pema, Monika ; Castelli, Maddalena ; Casanova, Ariel E. ; Mollica, Luca ; Banzi, Manuela ; Boca, Manila ; Antignac, Corinne ; Saunier, Sophie ; Musco, Giovanna ; Boletta, Alessandra. / Nephrocystin-1 forms a complex with polycystin-1 via a polyproline Motif/SH3 domain interaction and regulates the apoptotic response in mammals. In: PLoS One. 2010 ; Vol. 5, No. 9.
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abstract = "Mutations in PKD1, the gene encoding for the receptor Polycystin-1 (PC-1), cause autosomal dominant polycystic kidney disease (ADPKD). The cytoplasmic C-terminus of PC-1 contains a coiled-coil domain that mediates an interaction with the PKD2 gene product, Polycystin-2 (PC-2). Here we identify a novel domain in the PC-1 C-terminal tail, a polyproline motif mediating an interaction with Src homology domain 3 (SH3). A screen for interactions using the PC-1 C-terminal tail identified the SH3 domain of nephrocystin-1 (NPHP1) as a potential binding partner of PC-1. NPHP1 is the product of a gene that is mutated in a different form of renal cystic disease, nephronophthisis (NPHP). We show that in vitro pull-down assays and NMR structural studies confirmed the interaction between the PC-1 polyproline motif and the NPHP1 SH3 domain. Furthermore, the two full-length proteins interact through these domains; using a recently generated model system allowing us to track endogenous PC-1, we confirm the interaction between the endogenous proteins. Finally, we show that NPHP1 trafficking to cilia does not require PC-1 and that PC-1 may require NPHP1 to regulate resistance to apoptosis, but not to regulate cell cycle progression. In line with this, we find high levels of apoptosis in renal specimens of NPHP patients. Our data uncover a link between two different ciliopathies, ADPKD and NPHP, supporting the notion that common pathogenetic defects, possibly involving de-regulated apoptosis, underlie renal cyst formation.",
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AU - Rowe, Isaline

AU - Gaetani, Massimiliano

AU - Bricoli, Barbara

AU - Muorah, Mordi

AU - Spitaleri, Andrea

AU - Mannella, Valeria

AU - Ricchiuto, Piero

AU - Pema, Monika

AU - Castelli, Maddalena

AU - Casanova, Ariel E.

AU - Mollica, Luca

AU - Banzi, Manuela

AU - Boca, Manila

AU - Antignac, Corinne

AU - Saunier, Sophie

AU - Musco, Giovanna

AU - Boletta, Alessandra

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N2 - Mutations in PKD1, the gene encoding for the receptor Polycystin-1 (PC-1), cause autosomal dominant polycystic kidney disease (ADPKD). The cytoplasmic C-terminus of PC-1 contains a coiled-coil domain that mediates an interaction with the PKD2 gene product, Polycystin-2 (PC-2). Here we identify a novel domain in the PC-1 C-terminal tail, a polyproline motif mediating an interaction with Src homology domain 3 (SH3). A screen for interactions using the PC-1 C-terminal tail identified the SH3 domain of nephrocystin-1 (NPHP1) as a potential binding partner of PC-1. NPHP1 is the product of a gene that is mutated in a different form of renal cystic disease, nephronophthisis (NPHP). We show that in vitro pull-down assays and NMR structural studies confirmed the interaction between the PC-1 polyproline motif and the NPHP1 SH3 domain. Furthermore, the two full-length proteins interact through these domains; using a recently generated model system allowing us to track endogenous PC-1, we confirm the interaction between the endogenous proteins. Finally, we show that NPHP1 trafficking to cilia does not require PC-1 and that PC-1 may require NPHP1 to regulate resistance to apoptosis, but not to regulate cell cycle progression. In line with this, we find high levels of apoptosis in renal specimens of NPHP patients. Our data uncover a link between two different ciliopathies, ADPKD and NPHP, supporting the notion that common pathogenetic defects, possibly involving de-regulated apoptosis, underlie renal cyst formation.

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