Functional impairment of p16INK4A due to CDKN2A p.Gly23Asp missense mutation

Maria Chiara Scaini, Elisabetta Rossi, Paula Lobao Antunes de Siqueira Torres, Daniela Zullato, Monia Callegaro, Cinzia Casella, Monica Quaggio, Simona Agata, Sandro Malacrida, Vanna Chiarion-Sileni, Antonella Vecchiato, Mauro Alaibac, Marco Montagna, Graham J. Mann, Chiara Menin, Emma D'Andrea

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The CDKN2A locus encodes for two distinct tumor suppressor proteins, p16INK4A and p14ARF, involved in cell cycle regulation. CDKN2A germline mutations have been associated with familial predisposition to melanoma and other tumor types. Besides bona-fide pathogenic mutations, many sequence variants have been identified, but their effect is not well known. We detected the p.Gly23Asp missense mutation in one of the two tested melanoma patients of a family with three melanoma cases. Even though the mutated amino acid is located in a conserved domain that specifically binds to and blocks the function of CDK4/6, its lack of segregation with disease suggested a series of functional assays to discriminate between a pathogenic variant and a neutral polymorphism. The effect of this mutation has been investigated exploiting four p16INK4A properties: its ability (i) to bind CDK4, (ii) to inhibit pRb phosphorylation, (iii) to evenly localize in the cell, and (iv) to cause cell cycle arrest. The mutant protein properties were evaluated transfecting three different cell lines (U2-OS and NM-39, both p16-null, and SaOS 2, p53 and pRb-null) with plasmids expressing either p16wt, p1623Asp, or the p1632Pro pathogenic variant. We found that p1623Asp was less efficient than p16wt in CDK4 binding, in inhibiting pRb phosphorylation, in inducing G1 cell cycle arrest; moreover, its pattern of distribution throughout the cell was suggestive of protein aggregation, thus assessing a pathogenic role for p1623Asp in familial melanoma.

Original languageEnglish
Pages (from-to)26-32
Number of pages7
JournalMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
Volume671
Issue number1-2
DOIs
Publication statusPublished - Dec 1 2009

Fingerprint

Missense Mutation
Melanoma
Tumor Suppressor Protein p14ARF
Phosphorylation
G1 Phase Cell Cycle Checkpoints
Mutation
Aptitude
Germ-Line Mutation
Mutant Proteins
Cell Cycle Checkpoints
Cell Cycle
Plasmids
Amino Acids
Cell Line
Neoplasms
Proteins

Keywords

  • CDKN2A locus
  • Familial melanoma
  • Functional assays
  • p16 unclassified variant

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

Cite this

Functional impairment of p16INK4A due to CDKN2A p.Gly23Asp missense mutation. / Scaini, Maria Chiara; Rossi, Elisabetta; de Siqueira Torres, Paula Lobao Antunes; Zullato, Daniela; Callegaro, Monia; Casella, Cinzia; Quaggio, Monica; Agata, Simona; Malacrida, Sandro; Chiarion-Sileni, Vanna; Vecchiato, Antonella; Alaibac, Mauro; Montagna, Marco; Mann, Graham J.; Menin, Chiara; D'Andrea, Emma.

In: Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis, Vol. 671, No. 1-2, 01.12.2009, p. 26-32.

Research output: Contribution to journalArticle

Scaini, Maria Chiara ; Rossi, Elisabetta ; de Siqueira Torres, Paula Lobao Antunes ; Zullato, Daniela ; Callegaro, Monia ; Casella, Cinzia ; Quaggio, Monica ; Agata, Simona ; Malacrida, Sandro ; Chiarion-Sileni, Vanna ; Vecchiato, Antonella ; Alaibac, Mauro ; Montagna, Marco ; Mann, Graham J. ; Menin, Chiara ; D'Andrea, Emma. / Functional impairment of p16INK4A due to CDKN2A p.Gly23Asp missense mutation. In: Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis. 2009 ; Vol. 671, No. 1-2. pp. 26-32.
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AU - Rossi, Elisabetta

AU - de Siqueira Torres, Paula Lobao Antunes

AU - Zullato, Daniela

AU - Callegaro, Monia

AU - Casella, Cinzia

AU - Quaggio, Monica

AU - Agata, Simona

AU - Malacrida, Sandro

AU - Chiarion-Sileni, Vanna

AU - Vecchiato, Antonella

AU - Alaibac, Mauro

AU - Montagna, Marco

AU - Mann, Graham J.

AU - Menin, Chiara

AU - D'Andrea, Emma

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