Temperature-sensitive RB mutations linked to incomplete penetrance of familial retinoblastoma in 12 families

Am J Hum Genet. 1999 Oct;65(4):1040-6. doi: 10.1086/302581.


The tumor-suppressor activity of the retinoblastoma protein (RB) is encoded within a protein-binding ("pocket") domain that is targeted for mutations in all cases of familial retinoblastoma and in many common adult cancers. Although familial retinoblastoma is a paradigm for a highly penetrant, recessive model of tumorigenesis, the molecular basis for the phenotype of incomplete penetrance of familial retinoblastoma is undefined. We studied the RB pocket-binding properties of three independent, mutant RB alleles that are present in the germline of 12 kindreds with the phenotype of incomplete penetrance of familial retinoblastoma. Each arises from alterations of single codons within the RB pocket domain (designated "delta 480," "661W," or "712R"). Under the same conditions, we studied the properties of wild-type (WT) RB, an RB point mutant isolated from a lung carcinoma sample (706F) and an adjacent, in vitro-generated point mutant (707W). The delta 480, 661W, and 712R mutants lack pocket protein-binding activity in vitro but retain the WT ability to undergo cyclin-mediated phosphorylation in vivo. Each of the low-penetrant RB mutants exhibits marked enhancement of pocket protein binding when the cells are grown at reduced temperature. In contrast, in this temperature range, no change in binding activity is seen with WT RB, the 706F mutant, or the 707W mutant. We have demonstrated that many families with incomplete penetrance of familial retinoblastoma carry unstable, mutant RB alleles with temperature-sensitive pocket protein-binding activity. The variable frequency for tumor development in these families may result from reversible fluctuations in a threshold level of RB pocket-binding activity.

MeSH terms

  • Alleles
  • Amino Acid Substitution / genetics
  • Binding Sites
  • Cyclins / metabolism
  • Genes, Recessive / genetics
  • Genes, Retinoblastoma / genetics*
  • Germ-Line Mutation / genetics
  • Humans
  • Lung Neoplasms / genetics
  • Molecular Sequence Data
  • Mutation, Missense / genetics
  • Penetrance*
  • Phosphorylation
  • Point Mutation / genetics*
  • Protein Binding
  • Protein Structure, Tertiary
  • Retinoblastoma / genetics*
  • Retinoblastoma / metabolism
  • Retinoblastoma Protein / chemistry
  • Retinoblastoma Protein / genetics*
  • Retinoblastoma Protein / metabolism
  • Temperature
  • Two-Hybrid System Techniques


  • Cyclins
  • Retinoblastoma Protein

Associated data

  • OMIM/180200