TY - JOUR
T1 - Interaction of the single-stranded DNA-binding protein Purα with the human polyomavirus JC virus early protein T-antigen
AU - Gallia, Gary L.
AU - Safak, Mahmut
AU - Khalili, Kamel
PY - 1998/12/4
Y1 - 1998/12/4
N2 - Large T-antigen, the major regulatory protein encoded by polyomaviruses, including Simian Virus 40 (SV40) and JC virus (JCV), is a multifunctional phosphoprotein that is involved in many viral and cellular events. In addition to its integral role in viral replication and cellular transformation, T-antigen also regulates transcription of both viral and cellular genes. In particular, the viral late promoter has been used as a model for the analysis of T-antigen-mediated transcriptional activation. Earlier studies have demonstrated that the cellular protein Purα is able to attenuate the transcriptional activity of JCV T-antigen. We investigated the mechanism whereby Purα affects T-antigen function. Co-immunoprecipitation studies demonstrated that Purα and JCV T-antigen associate in vivo, and glutathione S-transferase affinity binding assays revealed that these two proteins interact in vitro. Moreover, we localized the sequences of Purα that are important for the interaction between Purα and JCV T-antigen. In addition, we demonstrated that Purα interacts with the SV40 T-antigen. Transient transfection studies demonstrated that Purα and JCV T-antigen interact functionally as well. More specifically, Purα and a deletion mutant that interacts with T-antigen attenuated T-antigen-mediated transcriptional activation. A Purα deletion mutant that is unable to interact with JCV T- antigen, however, was found to be incapable of abrogating JCV T-antigen transactivation. Taken together, these data demonstrate that Purα and T- antigen interact both physically and functionally and that this interaction modulates T-antigen-mediated transcriptional activation. The implication of these findings with respect to the cellular role of Purα is discussed.
AB - Large T-antigen, the major regulatory protein encoded by polyomaviruses, including Simian Virus 40 (SV40) and JC virus (JCV), is a multifunctional phosphoprotein that is involved in many viral and cellular events. In addition to its integral role in viral replication and cellular transformation, T-antigen also regulates transcription of both viral and cellular genes. In particular, the viral late promoter has been used as a model for the analysis of T-antigen-mediated transcriptional activation. Earlier studies have demonstrated that the cellular protein Purα is able to attenuate the transcriptional activity of JCV T-antigen. We investigated the mechanism whereby Purα affects T-antigen function. Co-immunoprecipitation studies demonstrated that Purα and JCV T-antigen associate in vivo, and glutathione S-transferase affinity binding assays revealed that these two proteins interact in vitro. Moreover, we localized the sequences of Purα that are important for the interaction between Purα and JCV T-antigen. In addition, we demonstrated that Purα interacts with the SV40 T-antigen. Transient transfection studies demonstrated that Purα and JCV T-antigen interact functionally as well. More specifically, Purα and a deletion mutant that interacts with T-antigen attenuated T-antigen-mediated transcriptional activation. A Purα deletion mutant that is unable to interact with JCV T- antigen, however, was found to be incapable of abrogating JCV T-antigen transactivation. Taken together, these data demonstrate that Purα and T- antigen interact both physically and functionally and that this interaction modulates T-antigen-mediated transcriptional activation. The implication of these findings with respect to the cellular role of Purα is discussed.
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U2 - 10.1074/jbc.273.49.32662
DO - 10.1074/jbc.273.49.32662
M3 - Article
C2 - 9830007
AN - SCOPUS:0032484017
SN - 0021-9258
VL - 273
SP - 32662
EP - 32669
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 49
ER -