Mutational and functional analysis of the tumor-suppressor PTPRD in human melanoma

Hum Mutat. 2014 Nov;35(11):1301-10. doi: 10.1002/humu.22630. Epub 2014 Sep 10.

Abstract

Protein tyrosine phosphatases (PTPs) tightly regulate tyrosine phosphorylation essential for cell growth, adhesion, migration, and survival. We performed a mutational analysis of the PTP gene family in cutaneous metastatic melanoma and identified 23 phosphatase genes harboring somatic mutations. Among these, receptor-type tyrosine-protein phosphatase delta (PTPRD) was one of the most highly mutated genes, harboring 17 somatic mutations in 79 samples, a prevalence of 21.5%. Functional evaluation of six PTPRD mutations revealed enhanced anchorage-dependent and anchorage-independent growth. Interestingly, melanoma cells expressing mutant PTPRD were significantly more migratory than cells expressing wild-type PTPRD or vector alone, indicating a novel gain-of-function associated with mutant PTPRD. To understand the molecular mechanisms of PTPRD mutations, we searched for its binding partners by converting the active PTPRD enzyme into a "substrate trap" form. Using mass spectrometry and coimmunoprecipitation, we report desmoplakin, a desmosomal protein that is implicated in cell-cell adhesion, as a novel PTPRD substrate. Further analysis showed reduced phosphatase activity of mutant PTPRD against desmoplakin. Our findings identify an essential signaling cascade that is disrupted in melanoma. Moreover, because PTPRD is also mutated in glioblastomas and adenocarcinoma of the colon and lung, our data might be applicable to a large number of human cancers.

Keywords: PTPRD; cell migration; desmoplakin; desmosomes; somatic mutations.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation
  • DNA Mutational Analysis
  • Desmoplakins / metabolism
  • Gene Expression
  • Humans
  • Intercellular Junctions / metabolism
  • Melanoma / genetics*
  • Melanoma / metabolism*
  • Models, Biological
  • Mutation*
  • Phosphorylation
  • Protein Binding
  • Protein Transport
  • Receptor-Like Protein Tyrosine Phosphatases, Class 2 / genetics*
  • Receptor-Like Protein Tyrosine Phosphatases, Class 2 / metabolism*
  • Substrate Specificity

Substances

  • Carrier Proteins
  • Desmoplakins
  • PTPRD protein, human
  • Receptor-Like Protein Tyrosine Phosphatases, Class 2