Use of sequential immunoprecipitation to reveal discrete, separable populations of SV40 T-antigen binding to host cellular proteins.
Sequential immunoprecipitations were carried out to determine the usefulness of this method for separating subpopulations of SV40 T-antigen complexed to various combinations of the cellular growth regulatory proteins pRB, p107, and p53. This approach was used successfully to separate discrete populations of SV40 T-antigen in a quatramolecular complex with pRB, p53, and p107, a trimolecular complex with pRB and p107, and a trimolecular complex with p107 and p53. This method was used as the first step towards isolating T-antigen for subsequent phosphopeptide mapping to address whether alterations in the overt phosphorylation of this viral oncoprotein is a major determinating factor to separation of T-antigen populations by complexing with different combinations of cellular growth regulatory proteins.
Ludlow
J W
JW
University of Rochester Cancer Center, Division of Developmental Therapeutics, NY 14642, USA. jludlow@cc.urmc.rochester.edu
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D009687:Nuclear Proteins / Q000276:immunology*; D011233:Precipitin Tests / Q000379:methods*; D016160:Retinoblastoma Protein / Q000276:immunology*; D013539:Simian virus 40 / Q000235:genetics / Q000276:immunology*; D016159:Tumor Suppressor Protein p53 / Q000276:immunology*
D000952:Antigens, Polyomavirus Transforming; D009687:Nuclear Proteins; D016160:Retinoblastoma Protein; D016159:Tumor Suppressor Protein p53