Mutation of the LXCXE binding cleft of pRb facilitates transformation by ras in vitro but does not promote tumorigenesis in vivo

PLoS One. 2013 Aug 6;8(8):e72236. doi: 10.1371/journal.pone.0072236. Print 2013.

Abstract

Background: The Retinoblastoma protein (pRB) is a key tumor suppressor that is functionally inactivated in most cancers. pRB regulates the cell division cycle and cell cycle exit through protein-protein interactions mediated by its multiple binding interfaces. The LXCXE binding cleft region of pRB mediates interactions with cellular proteins that have chromatin regulatory functions. Chromatin regulation mediated by pRB is required for a stress responsive cell cycle arrest, including oncogene induced senescence. The in vivo role of chromatin regulation by pRB during senescence, and its relevance to cancer is not clear.

Methodology/principal findings: Using gene-targeted mice, uniquely defective for pRB mediated chromatin regulation, we investigated its role during transformation and tumor progression in response to activation of oncogenic ras. We report that the pRB(∆L) mutation confers susceptibility to escape from HrasV12 induced senescence and allows transformation in vitro, although these cells possess high levels of DNA damage. Intriguingly, LSL-Kras, Rb1 (∆L/∆L) mice show delayed lung tumor formation compared to controls. This is likely due to the increased apoptosis seen in the early hyperplastic lesions shortly following ras activation that inhibits tumor progression. Furthermore, DMBA treatment to induce sporadic ras mutations in other tissues also failed to reveal greater susceptibility to cancer in Rb1 (∆L/∆L) mice.

Conclusions/significance: Our data suggests that chromatin regulation by pRB can function to limit proliferation, but its loss fails to contribute to cancer susceptibility in ras driven tumor models because of elevated levels of DNA damage and apoptosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Binding Sites
  • Carcinogenesis*
  • Cell Proliferation
  • Cell Transformation, Neoplastic / genetics*
  • Cellular Senescence
  • Chromatin / genetics
  • Chromatin / metabolism
  • Clone Cells / metabolism
  • Clone Cells / pathology
  • DNA Damage / genetics
  • Gene Expression Regulation
  • Hyperplasia / genetics
  • Hyperplasia / metabolism
  • Hyperplasia / pathology
  • Lung / pathology
  • Lung Neoplasms / genetics
  • Lung Neoplasms / metabolism
  • Lung Neoplasms / pathology
  • Mice
  • Mutation*
  • Oncogenes / genetics
  • Proto-Oncogene Proteins p21(ras) / genetics*
  • Proto-Oncogene Proteins p21(ras) / metabolism*
  • Retinoblastoma Protein / chemistry
  • Retinoblastoma Protein / genetics*
  • Retinoblastoma Protein / metabolism*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Chromatin
  • Retinoblastoma Protein
  • Tumor Suppressor Protein p53
  • Hras protein, mouse
  • Proto-Oncogene Proteins p21(ras)

Grants and funding

This work was supported by the Canadian Cancer Society Research Institute, grant number 700720. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.