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Isolation of the Rb-related p130 through its interaction with CDK2 and cyclins

G J Hannon1, D Demetrick, D Beach

  • 1Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, New York 11724.

Genes & Development
|December 1, 1993
PubMed

Insights

Researchers identified Rbr-2, a novel retinoblastoma gene family member, interacting with human CDK2 via a yeast cyclin bridge. This protein, p130Rbr-2, binds viral oncoproteins and D-type cyclins, suggesting a role in tumor development.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Virology

Background:

  • The retinoblastoma (Rb) protein family plays a crucial role in cell cycle regulation.
  • Understanding interactions with cyclin-dependent kinases (CDKs) like CDK2 is vital for cancer research.
  • Viral oncoproteins often target Rb family proteins to disrupt cell cycle control.

Purpose of the Study:

  • To identify novel human proteins interacting with CDK2 using a yeast two-hybrid system.
  • To characterize the newly identified Rbr-2 protein and its relationship to known Rb family members.
  • To investigate the interaction of Rbr-2 with viral oncoproteins and cell cycle regulators.

Main Methods:

  • Yeast two-hybrid screening to identify protein interactions.
  • Sequence analysis to determine protein homology and identify Rbr-2.
  • In vitro binding assays to study protein-protein interactions, including with cyclins and viral oncoproteins.
  • Gene mapping to determine the chromosomal location of the Rbr-2 gene.

Main Results:

  • A novel retinoblastoma gene family member, Rbr-2 (Rb-related), was isolated.
  • Rbr-2 protein shares homology with Rb and p107, particularly in the pocket domain, and is identified as p130.
  • p130Rbr-2 binds to SV40 large T antigen and adenovirus E1A via its pocket domain.
  • p130Rbr-2 interacts with D-type cyclins but not directly with CDK2 in vitro.
  • The gene encoding p130Rbr-2 maps to chromosome 16q13, a region associated with genomic alterations in tumors.

Conclusions:

  • The study identified p130Rbr-2 as a novel Rb family member interacting with viral oncoproteins and D-type cyclins.
  • The interaction between CDK2 and p130Rbr-2 in yeast is likely mediated by a yeast-specific cyclin.
  • The chromosomal location of p130Rbr-2 suggests its potential involvement in human tumorigenesis.

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