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. 1993 Apr;67(4):2235–2244. doi: 10.1128/jvi.67.4.2235-2244.1993

Novel monoclonal antibodies that differentiate between the binding of pp60c-src or protein phosphatase 2A by polyomavirus middle T antigen.

S M Dilworth 1, V P Horner 1
PMCID: PMC240352  PMID: 7680389

Abstract

Fourteen pGEX plasmids that express defined regions of polyomavirus middle T antigen in bacteria have been constructed. These polypeptides have been used to generate 18 new monoclonal antibodies directed against the unique portion of middle T and to map the approximate position of the antibody recognition sites onto the protein sequence. All of the antibodies effectively immunoprecipitate middle T and the associated 60- and 35-kDa components of protein phosphatase 2A. Four of the antibodies, however, do not react with middle T when it is bound to pp60c-src. These four probably bind to amino acids 203 to 218 of the middle T protein sequence, which are encoded by the mRNA immediately 3' to the splice junction that creates the C-terminal unique region. This suggests that additional middle T sequences are required for middle T's interaction with pp60c-src than are needed for its binding to protein phosphatase 2A. The antibodies localize this extra region and provide a means of distinguishing between these two associations.

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Selected References

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